HITTA DENNAUS UNUTRITION 20180042989A1 ADULT CLIO III ( 19) United States (12 ) Patent Application Publication (10 ) Pub. No. : US 2018/ 0042989 A1 Aversa et al. (43 ) Pub . Date : Feb . 15 , 2018 ( 54 ) IMMUNOMODULATORY COMPOSITIONS Publication Classification (51 ) Int. Cl. ( 71 ) Applicant: Sigmoid Pharma Limited , Dublin ( IE ) A61K 38 / 13 ( 2006 .01 ) (72 ) Inventors : Vincenzo Aversa , Ridgewood Swords A61K 31/ 502 ( 2006 . 01) ( IE ); Ivan Coulter, Mount Merrion A61K 31/ 436 (2006 . 01) ( IE ) ; Mónica Torres Rosa , Dublin (IE ) ; A61K 45 / 06 ( 2006 .01 ) Bernard Francis McDonald , A61K 9 / 16 (2006 .01 ) Castleblayney (IE ) A61K 9 /50 ( 2006 .01 ) (52 ) U . S . CI. (73 ) Assignee : Sigmoid Pharma Limited , Dublin ( IE ) CPC ...... A61K 38 / 13 ( 2013 .01 ) ; A6IK 9 / 167 (2013 .01 ) ; A61K 9 / 16 (2013 . 01 ) ; A61K 9 /5036 (21 ) Appl . No. : 15 /791 , 104 ( 2013 .01 ) ; A61K 31 / 436 ( 2013 .01 ) ; A61K 9 /5084 ( 2013 .01 ); A61K 9 / 1658 ( 2013 .01 ) ; (22 ) Filed : Oct . 23, 2017 A61K 45 / 06 ( 2013 .01 ) ; A61K 31/ 502 (2013 . 01 ); A61K 9 /5042 (2013 . 01 ) ; A61K Related U . S . Application Data 2300 / 00 (2013 .01 ) (63 ) Continuation of application No . 13 /989 ,372 , filed on May 23 , 2013 , now Pat . No . 9 ,821 ,024 , filed as (57 ) ABSTRACT application No. PCT /EP2011 /071088 on Nov . 25 , 2011. Immunomodulator formulations for use in the treatment of disease of the GI tract. The formulations comprise a Foreign Application Priority Data hydroxylase inhibitor and /or an immunosuppressant. Exem (30 ) plary formulations comprise hydralazine as a hydroxylase Nov . 25, 2010 (GB ) ...... 1020032 .7 inhibitor and / or cyclosporin A as an immunosuppressant. Patent Application Publication Feb . 15, 2018 Sheet 1 of 43 US 2018 / 0042989 A1
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IMMUNOMODULATORY COMPOSITIONS nant ( IBS- C ) and irritable bowel syndrome mixed ( IBS M ) ; diverticulosis , diverticulitis and endometriosis . CROSS REFERENCE TO RELATED [0007 ] Extraintestinal disorders in which a leaky intestinal APPLICATIONS epithelial barrier have been implicated include rheumatic [ 0001] This is a continuation of U . S . application Ser. No. disorders, rheumatoid arthritis, temporomandibular joint 13 / 989 ,372 , filed on May 23 , 2013 , which is the U . S . syndrome, type 1 diabetes , multiple sclerosis , atopic derma National Stage of International Application No . PCT/ titis , psoriasis , a chronic pain syndrome, fibromyalgia , EP2011/ 071088 , filed Nov. 25 , 2011 , which was published chronic fatigue syndrome, depressive disorders , affective in English under PCT Article 21( 2 ) , which in turn claims the disorders and attention disorders, colorectal carcinoma, benefit of Great Britain Application No. 1020032 . 7 , filed adenocarcinoma, and chronic heart failure . Nov . 25 , 2010 , all of which are incorporated herein in their entirety . [0008 ] For example , the reader is referred to Vaarala O , Atkinson MA , Neu J ( October 2008 ) . The “ perfect storm ” for type 1 diabetes: the complex interplay between intestinal FIELD microbiota , gut permeability , and mucosal immunity '. Dia [0002 ] This invention relates to immunomodulatory com betes 57 ( 10 ): 2555 -62 ; Liu Z , Li N , Neu J ( April 2005 ). positions and to therapeutic methods using immunomodu “ Tight junctions, leaky intestines, and pediatric diseases” . latory compositions. The invention also relates to methods Acta Paediatr . 94 ( 4 ) : 386 - 93 ; Maes M . Inflammatory and of making the compositions, methods of using them , and oxidative and nitrosative stress pathways underpinning other subject matter. chronic fatigue , somatization and psychosomatic symptoms. Curr Opin Psychiatry . 2009 January ; 22 ( 1 ) : 75 - 83 ; de Mag BACKGROUND istris L , Familiari V , Pascotto A , Sapone A , Frolli A , lardino [0003 ] Disorders of the intestine , disorders of the colon , P , Carteni M , De Rosa M , Francavilla R , Riegler G , Militerni and disorders associated with or suspected of being associ R , Bravaccio C . Alterations of the intestinal barrier in ated with a dysfunctional intestine and /or dysfunctional patients with autism spectrum disorders and in their first colon , are an unmet medical need . The present invention degree relatives . J Pediatr GastroenterolNutr . 2010 October ; comprises products , methods and uses which are useful in 51( 4 ) : 418 - 24 ; Sandek A , Rauchhaus M , Anker S D , von relation to treating such disorders. Haehling S (September 2008 ). “ The emerging role of the gut [ 0004 ] More particularly , inflammatory Bowel Disease in chronic heart failure " . Curr Opin Clin Nutr Metab Care 11 (IBD ) , which includes ulcerative colitis and Crohn ' s Dis ( 5 ): 632 - 9 ; Terjung B , Spengler U (February 2009 ). “ Atypi ease , is a common disease with severe morbidity and an cal p - ANCA in PSC and AIH : a hint toward a “ leaky gut” ? ” . extremely limited therapeutic repertoire . In most cases , Clin Rev Allergy Immunol 36 ( 1 ) : 40 -51 ; Intestinal mucosal surgical intervention is necessary due to the failure of permeability in inflammatory rheumatic diseases . II . Role of successful pharmacologic intervention . Improved therapy disease . Mielants H , De Vox M , Goemaere S , Schelstraete K , for IBD is therefore extremely desirable . Cuvelier C , Goethals K , Maertens M , Ackerman C , Veys E [0005 ] Current pharmacologic therapy for IBD is limited M , J Rheumatol. 1991 March ; 18 ( 3 ) : 394 -400 ; Intestinal to steroids , sulfasalizine and TNF antibodies all of which mucosal permeability in inflammatory rheumatic diseases. I. have significant limitations due to side effects caused by the Role of antiinflammatory drugs . Mielants H , Goemaere S , high levels of systemic administration thatmust be given to De Vos M , Schelstraete K , Goethals K , Maertens M , Ack achieve therapeutic effects . The underlying causes of inflam erman C , Veys E M . J Rheumatol. 1991 March ; 18 ( 3 ) : 389 matory bowel disease are not clearly understood but the 93 ; Intestinal permeability and atopic disease . Mackie R M . symptoms have been treated through the use of anti - inflam Lancet. 1981 Jul . 18 ; 2 (8238 ): 155 . matory agents that often are not specific and lead to serious [0009 ] The reader is referred to a recent review by Andy side effects , especially if administered chronically . In addi Wullaert , Marion C Bonnet and Manolis Pasparakis in Cell tion to being toxic , most current anti - inflammatory thera Research (2011 ) 21 : 146 - 158 for an understanding of the role peutics have limited solubility , permeability or stability of NF- kB in the regulation of epithelial homeostasis and resulting in high drug doses, frequent administration or inflammation , and this publication and all the references administered by injection or suppositories , thus therapeutic mentioned in it are included herein by reference in their options in IBD are very limited . Hydroxylase inhibitors have entirety . In summary , a number of in vivo studies in genetic a potential role in the treatment of IBD . See The Hydroxy mouse models over the past years have revealed that NF - KB lase Inhibitor Dimethyloxalylglycine Is Protective in a inhibition can also trigger chronic inflammatory conditions. Murine Model of Colitis Gastroenterology , Volume 134 , This function of NF -kB appears to be particularly important Issue 1 , January 2008 , Pages 156 - 165. el. Eoin P . Cummins, at epithelial surfaces , where NF- kB activity in epithelial Fergal Seeballuck , Stephen J . Keely, Niamh E . Mangan , cells is required for the maintenance of immune homeosta John J . Callanan , Padraic G . Fallon , Cormac T . Taylor. sis . Therefore , proper regulation of NF -kB activation at [0006 ] A permeable “ leaky” ) intestinal epithelial barrier epithelial interfaces is crucial for the maintenance of physi has been implicated in intestinal and extraintestinal diseases . ological tissue homeostasis and for efficient host defense Examples of such diseases are the intestinal diseases inflam against environmental insults . NF -kB inhibition sensitizes matory bowel disease , celiac disease , Crohn ' s disease, ulcer epithelial cells to stress - inducing stimuli coming either from ative colitis , GI- GVHD , gastroenteritis , duodenitis , jejunitis , the environment ( e . g . , microorganisms) or from immune ileitis , peptic ulcer, Curling ' s ulcer , appendicitis , colitis , cells ( e . g . , cytokines ) and compromises their viability result pseudomembraneous colitis , irritable bowel syndrome ing in the deregulation of tissue immune homeostasis and including Irritable bowel syndrome diarrhea predominant triggering inflammation . Moreover , NF - kB is known to ( IBS - D ) , irritable bowel syndrome constipation predomi protect cells from a wide variety of cell death triggers. US 2018 / 0042989 A1 Feb . 15 , 2018
[0010 ] A number of studies have showed that inhibition of [0014 ] Included in the invention are pharmaceutical com NF -kB activation specifically in the intestinal epithelium positions wherein at least some of the minibeads have a causes severe intestinal inflammation . Thus , mice lacking controlled release coating adapted for the coated minibeads NF -kB signaling protein NEMO specifically in intestinal to release hydralazine and cyclosporin in the colon . The epithelial cells (IECs ) developed severe chronic colitis char coated minibeads may be coated with a coating comprising acterized by epithelial ulceration , elevated expression of a pH independent polymer and a polymer specifically sus proinflammatory mediators and infiltration of immune cells . ceptible of degradation by bacterial enzymes in the colon . Complete abrogation of canonical NF -KB activity in the The pH independent polymer may be ethylcellulose and the intestinal epithelium , achieved by ablation of NEMO (or by polymer specifically susceptible of degradation by bacterial combined deficiency of both IKK1 and IKK2 ) , caused enzymes in the colon may be pectin . severe colon inflammation , demonstrating that IKK /NF -KB [0015 ] The pharmaceutical composition may be for use in signaling performs essential homeostasis - preserving func a therapy selected from : maintaining the health of the tions in the colonic epithelium . Patients carrying hypomor gastrointestinal tract, restoring or improving the health of phic mutations in NEMO usually suffer from severe immu the gastrointestinal tract and delaying the progression of a nodeficiency and developmental skin defects , but some of disorder of the gastrointestinal tract . The pharmaceutical these patients also develop colitis . Interestingly , hematopoi composition my be for use in treating , or delaying the etic stem cell transplantation (HSCT ) is effective in treating progression of, a disorder selected from intestinal disorders the immunodeficiency, but does not improve the colitis which are inflammatory and / or fibrotic , or is for use in phenotype. On the contrary , HSCT often worsens pre maintenance therapy of a patient who has suffered from or existing colitis or even triggers colon inflammation in is suffering from such a disorder. The disorder may be patients who did not suffer from it before transplantation , selected from celiac disease , HIV or another enteropathy, suggesting that impaired NF - kB signaling in non - he pouchitis , cachexia , or the composition may be for use in matopoietic cells is responsible for colitis development. It preventing or reducing chemotherapy - induced or radiation would therefore be desirable to up -regulate NF -kB activity therapy - induced gastrointestinal insult and inflammatory in subjects who would benefit therefrom , for example in bowel disease , and combinations thereof. Celiac disease is patients who suffer from or have suffered from inflammatory primarily a disorder of the small intestine and, for treatment orders of the GI tract, for example those suffering from of this disease , formulations are suitably adapted to release graft -versus host disease following HSCT. active in the small intestine , e . g . minicapsules include an enteric barrier or coating for dissolution in the small intes BRIEF SUMMARY OF THE DISCLOSURE tine , for example in the duodenum . However, celiac disease [0011 ] This specification contains data indicating that may also manifest itself in the colon as microscopic colitis combination therapy with hydralazine and cyclosporin A , and IBD . Further, patients suffering from celiac disease particularly where the proportion of hydralazine is not too suffer an increased risk of also suffering from ulcerative low , has potential for prophylactic and therapeutic treatment colitis . It will therefore often be beneficial for celiac patients of disorders of the gastrointestinal tract. See in particular to have the actives described herein delivered to the colon as Examples 1 - 3 and 6 - 9 . In one aspect, therefore , the invention well as the small intestine . The invention therefore provides provides a product comprising hydralazine and cyclosporin combination formulations comprising hydralazine and Ain a weight ratio (hydralazine :cyclosporin ) of from 1 : 10 to cyclosporin A which are for use in delivery of the actives in 10 : 1 . The weight ratio may be : from 1 : 5 to 5 : 1 ; from 1 : 2 to the small intestine and for use in combination with a 5 : 1 ; from 1 : 2 to 2 : 1 ; from 0 . 5 : 1 to 5 : 1 ; or from 0 . 5 : 1 to 2 : 1 . combination formulation comprising hydralazine and The weight ratio may be from 0 . 8 : 1 to 5 : 1 , e . g . from 0 . 8 : 1 cyclosporin A for us in delivery of the actives in the colon . to 2 : 1 or from 0 . 8 : 1 to 1 . 5 : 1 . The weight ratio may be from The two different combination formulations may be admin 1 : 1 to 5 : 1 or from 1 : 1 to 2 : 1 , e . g . from 1 : 1 to 1 . 5 : 1 . The istered simultaneously , sequentially or separately . For weight ratio may be at least 1 . 1 : 1 , e . g . from 1 . 1 : 1 to 2 : 1 or example , a unit dosage , e .g . a capsule , may comprise a first from 1 . 1 : 1 to 1. 5 : 1 . population of hydralazine / cyclosporin minicapsules having [0012 ] The product may be a pharmaceutical composition a controlled release coating to release the active in the small for oral administration . Advantageously , the cyclosporin Ais intestine and a second population of hydralazine / cyclosporin dissolved . Cyclosporin A is poorly soluble in water and more minicapsules having a controlled release coating to release soluble in a hydrophobic environment; the pharmaceutical the actives in the colon . See below for more about coatings . composition may therefore comprise a hydrophobic phase in The second population may constitute no more than 50 % of which the cyclosporin is dissolved . The pharmaceutical minibeads in the unit dosage form , e . g . at least 10 % or at composition may be a multiple minibead composition least 20 % and no more than 30 % or 40 % , for example from wherein the hydralazine and cyclosporin A are contained in 20 % - 40 % or 10 % - 30 % . the minibeads , each minibead comprising a water -soluble [0016 ] It will be appreciated that the invention also polymer matrix material and , dispersed within the matrix includes an active agent selected from ( i ) hydralazine and material, the hydrophobic phase, the hydralazine being ( ii ) cyclosporin A for oral administration with the other of ( i ) comprised in the matrix material. The matrix material may and ( ii ) in a ratio as above . Also provided is an active agent comprise a hydrophilic surfactant having an HLB value of at selected from ( i ) hydralazine and ( ii ) cyclosporin A for use least 15 and the hydrophobic phase comprises a non -ionic in oral combination therapy with the other in a ratio as surfactant having an HLB value of at least 10 but less than above . The two actives may be administered simultaneously , that of the hydrophilic surfactant. sequentially or separately, e . g . each in a separate formulation [ 0013 ] The pharmaceutical composition may be adapted as described herein . The active agent may be for use in a for at least a portion of each of the hydralazine and the method of treatment described above in relation to the cyclosporin to be released in the colon . product of the first aspect of the invention or for use in a US 2018 / 0042989 A1 Feb . 15 , 2018 method of treatment described elsewhere herein . Each active composition wherein the immunosuppressant and the agentmay be comprised in a multiple minibead formulation hydroxylase inhibitor are contained in the minibeads, each as described elsewhere herein . minibead comprising a water- soluble polymer matrix mate [ 0017 ] This specification also contains data indicating that rial and , dispersed within the matrix material, the hydro combination therapy with an immunosuppressant and a phobic phase . The hydroxylase inhibitor may be hydralazine hydroxylase inhibitor has potential for the therapeutic and or another water soluble drug and be comprised in the matrix prophylactic treatment of fibrotic intestinal orders. See in material. The matrix material may comprise a hydrophilic particular examples 6 and 7 . In a second aspect, therefore , surfactant having an HLB value of at least 15 and the the invention provides a pharmaceutical composition com hydrophobic phase comprises a non - ionic surfactant having prising an immunosuppressant and a hydroxylase inhibitor an HLB value of at least 10 but less than that of the and for use in treating, or delaying the progression , of a hydrophilic surfactant. fibrotic intestinal disorder, or for use in maintenance therapypo [ 0022 ] The pharmaceutical composition of the second of a patient who has suffered from or is suffering from a aspect may be adapted for at least a portion of each of the fibrotic intestinal disorder . The disorder may be selected immunosuppressant and the hydroxylase inhibitor to be from celiac disease and HIV enteropathy. The disorder may released in the colon . be another enteropathy , e . g . one disclosed herein . [0023 ] In the multiple minibead compositions of the sec 10018 ] See above for a discussion about the treatment of ond aspect, at least some of the minibeads have a controlled celiac disease with hydralazine and cyclosporin A . That release coating adapted for the coated minibeads to release discussion applies mutatis mutandis to formulations com the hydroxylase inhibitor and the immunosuppressant in the prising a combination of an immunosuppressant and a colon . The coated minibeads may be coated with a coating hydroxylase inhibitor. HIV enterophathy is likely to occur comprising a pH independent polymer and a polymer spe widely throughout the intestinal tract and therapies will cifically susceptible of degradation by bacterial enzymes in typically involve simultaneous, separate or sequential the colon . The pH independent polymer may be ethylcellu administration of at least two combination formulations for lose and /or the polymer specifically susceptible of degrada use in releasing their actives in respective different parts of tion by bacterial enzymes in the colon may be pectin . the GI tract , e . g . at least in a part of the small intestine and [ 0024 ] The invention also includes an active agent at least in the colon . A multiple minibead unit dosage form , selected from ( i ) an immunosuppressant and ( ii ) a hydroxy e . g . capsule , may therefore include at least two populations lase inhibitor for administration with the other of ( i ) and ( ii ) of minibeads , each for use in releasing their contained and for use in treating , or delaying the progression , of a actives in a respective different part of the GI tract , e . g . at fibrotic intestinal disorder , or for use in maintenance therapy least a population to release the actives in a part of the small of a patient who has suffered from or is suffering from a intestine ( for example duodenum ) and a population to fibrotic intestinal disorder . Also provided is an active agent release the actives in the ileum and / or colon . See below for selected from ( i ) an immunosuppressant and ( ii ) a hydroxy more about targeted release and coatings therefor. lase inhibitor for use in oral combination therapy with the [ 0019 ). This paragraph applies to all aspects and imple other and for use in treating , or delaying the progression , of mentations of the disclosure. The immunosuppressant may a fibrotic intestinal disorder , or for use in maintenance be selected from , or comprise , cyclosporins , tacrolimus , therapy of a patient who has suffered from or is suffering sirolimus pimecrolimus , angiotensin II inhibitors ( e . g . Val from ( e . g . continues to suffer from ) a fibrotic intestinal sartan , Telmisartan , Losartan , Irbesatan , Azilsartan , Olme disorder. The two actives may be administered simultane sartan , Candesartan , Eprosartan ) and ACE inhibitors e . g . ously, sequentially or separately, e . g . each in a separate sulfhydryl- containing agents (e . g. Captopril, Zofenopril) , formulation as described herein . Each active agent may be dicarboxylate -containing agents ( e . g . Enalapril, Ramipril, comprised in a multiple minibead formulation as described Quinapril, Perindopril, Lisinopril , Benazepril , Imidapril , elsewhere herein . The active agent may be comprised in a Zofenopril, Trandolapril ), phosphate - containing agents ( e . g . pharmaceutical composition comprising the immunosup Fosinopril ) , casokinins, lactokinins and lactotripeptides. The pressant and the hydroxylase inhibitor and for use in treat hydroxylase inhibitor may be selected from , or comprise , ing , or delaying the progression , of a fibrotic intestinal DMOG , hydralazine , FG -4497 , FG4095 , AGN -2979 , disorder , or for use in maintenance therapy of a patient who metirosine , 3 - iodotyrosine , aquayamycin , bulbocapnine , has suffered from or is suffering from a fibrotic intestinal oudenone , TM 6008 , TM 6089 , siRNAs against hydroxy disorder. At least a portion of each of the immunosuppres lases and antisense therapeutics against hydroxylases , e . g . sant and the hydroxylase inhibitor may be released in the against PHD1, or a combination thereof. colon , each may therefore be included in a formulation 10020 ] In the second aspect, the immunosuppressant may adapted for at least a portion of its contained active to be be cyclosporin A and / or the hydroxylase inhibitor may be released in the colon ; suitably the formulation is a multiple hydralazine , e . g . it may be the case that the immunosup minibead formulation wherein at least some of the mini pressant is cyclosporin and the hydroxylase inhibitor is beads have a controlled release coating adapted for the hydralazine . The composition may comprise cyclosporin A coated minibeads to release their active agent in the colon ; and hydralazine in a hydralazine : cyclosporin weight ratio as such minibead formulations are described in more detail mentioned in relation to the first aspect, e. g . from 0 . 8 : 1 to elsewhere herein . 2 : 1 . 10025 ]. This specification contains data indicating that [ 0021] In the second aspect the pharmaceutical composi hydroxylase inhibitors are effective to up - regulate activity of tion may comprise a hydrophobic phase in which the immu NF -KB but in monotherapy may not be effective to bring nosuppressant is dissolved ; in particular the immunosup under control an existing inflammatory condition of the GI pressant may be a macrolide immunosuppressant. The tract. See in particular Example 8 . The data support a role for pharmaceutical composition may be a multiple minibead hydroxylase inhibitors in maintenance therapy , e . g . in sub US 2018 /0042989 A1 Feb . 15 , 2018 jects who previously had treatment to bring an inflammatory [ 0031 ] The invention therefore provides a method of treat condition of the GI tract under control and might benefit ing a warm - blooded animal e . g . a mammal, such as a human , from therapy , for example to maintain somewhere around to treat, or delay the progression of, a disorder selected from their existing level of GI tract health . Maintenance therapy intestinal disorders which are inflammatory and / or fibrotic , may control the symptoms of the disorder, e . g . stop , delay or or for use in maintenance therapy of an animal who has reduce a worsening of symptoms. Accordingly , in a third suffered from or is suffering from such a disorder, which aspect, there is provided a hydroxylase inhibitor for use in method comprises orally administering to said animal simul maintenance therapy of a patient who has suffered from or taneously , sequentially or separately hydralazine and is suffering from an inflammatory intestinal disorder . The cyclosporin A in a weight ratio (hydralazine :cyclosporin ) of disorder may be selected from (i ) inflammatory bowel from 1 : 10 to 10 : 1 e . g . 1 : 5 to 5 : 1 , for example 0 . 8 : 1 to 5 : 1 . disease and ( ii ) graft - versus -hose disease following Particular embodiments of this method comprise the admin hematopoietic stem cell transplantation . The disorder may istration of a composition according to any of the compo be colitis . sition embodiments described herein . The composition may [ 0026 ] The hydroxylase inhibitor may be selected from be administered in a therapeutically effective amount or in a those listed previously , e . g . it may be hydralazine . prophylactically effective amount. In particular, the disorder [0027 ] The hydroxylase inhibitor of the third aspect may is selected from celiac disease , HIV and other enteropathies be in solution phase in a pharmaceutical composition . The and inflammatory bowel disease , and combinations thereof. hydroxylase inhibitor may be sufficiently water - soluble for [0032 ] The invention therefore provides a method of treat adequate drug loading with the inhibitor in the solution ing a disorder selected from celiac disease , HIV enteropathy phase . The hydroxylase inhibitor may be sparingly soluble and inflammatory bowel disease , and combinations thereof, in water but more commonly is at least soluble and may be in a warm - blooded animal e . g . a mammal, such as a human , freely soluble or very soluble , as these terms are defined which method comprises orally administering to said animal herein . The pharmaceutical composition may be a multiple simultaneously , sequentially , or separately hydralazine and minibead composition , the minibeads comprising a water cyclosporin A in a weight ratio (hydralazine : cyclosporin ) of soluble polymer matrix in which a water hydroxylase inhibi from 1 : 10 to 10 : 1 e . g . 1 : 5 to 5 : 1 , for example 0 . 8 : 1 to 5 : 1 . tor is in solid solution . The minibeads may further comprise Particular embodiments of this method comprise the admin a hydrophobic phase dispersed in the matrix , the hydropho istration of a composition according to any of the compo bic phase being formed of material liquid at body tempera sition embodiments described herein . The composition may ture. The pharmaceutical composition may be adapted for at be administered in a therapeutically effective amount or in a least a portion of the hydroxylase inhibitor to be released in prophylactically effective amount. the colon . The hydroxylase inhibitor is not required to be in [0033 ] There is therefore provided a method of treating a the solution phase , e . g . it may be in the form of micro - or warm -blooded animal e. g . a mammal , such as a human , to nano - particles, in particular such a particulate hydroxylase treat, or delay the progression , of a fibrotic intestinal disor der, or for maintenance therapy of an animal which has inhibitor may be included in a water soluble polymer matrix suffered from or is suffering from a fibrotic intestinal dis if it is soluble , freely soluble or very soluble in water . order , which method comprises orally administering to said [ 0028 ] In the multiple minibead composition of the third animal simultaneously , sequentially or separately an immu aspect, at least some of the minibeads may have a controlled nosuppressant and a hydroxylase inhibitor. Particular release coating adapted for the coated minibeads to release embodiments of this method comprise the administration of hydralazine in the colon . The coated minibeads may be a composition according to any of the composition embodi coated with a coating comprising a pH independent polymer ments described herein . The composition may be adminis and a polymer specifically susceptible of degradation by tered in a therapeutically effective amount or in a prophy bacterial enzymes in the colon . The pH independent polymer lactically effective amount. In particular the disorder is may be ethylcellulose and /or the polymer specifically sus selected from celiac disease and HIV enteropathy. ceptible of degradation by bacterial enzymes in the colon [0034 ] The invention therefore provided a method of may be pectin . treating a disorder selected from celiac disease and HIV [ 0029 ] According to a further aspect of the invention , enteropathy in a warm - blooded animal e . g . a mammal, such there is provided a combination of actives or a composition as a human , which method comprises administering to said as defined herein for use in a method of treatment of the animal simultaneously , sequentially or separately an immu human or animal body by therapy or prophylaxis . nosuppressant and a hydroxylase inhibitor. Particular [0030 ] The invention therefore provides a method of treat embodiments of this method comprise the administration of ing a warm -blooded animal e . g . a mammal, such as a human , a composition according to any of the composition embodi to maintain the health of the gastrointestinal tract , to restore ments described herein . The composition may be adminis or improve the health of the gastrointestinal tract and / or to tered in a therapeutically effective amount or in a prophy delay the progression of a disorder of the gastrointestinal lactically effective amount. tract, which method comprises orally administering to said [0035 ] According to a further aspect of the invention , animal simultaneously , sequentially or separately hydrala there is provided a hydroxylase inhibitor as described herein zine and cyclosporin A in a weight ratio (hydralazine : for use in the method of treatment of the human or animal cyclosporin ) of from 1 : 10 to 10 : 1 , e . g . 1 : 5 to 5 : 1 , for body by therapy or prophylaxis . In particular the treatment example 0 .8 : 1 to 5 : 1 . Particular embodiments of this method is maintenance therapy . comprise the administration of a composition according to [0036 ] There is therefore provided a method for mainte any of the composition embodiments described herein . The nance therapy of a warm -blooded animal e . g . a mammal, composition may be administered in a therapeutically effec - such as a human , which has suffered from or is suffering tive amount or in a prophylactically effective amount. from an inflammatory intestinal disorder, which method US 2018 / 0042989 A1 Feb . 15 , 2018 comprises administered to said animal a hydroxylase inhibi tors and macrolide immunosuppressants; droplets of water tor. Particular embodiments of this method comprise the immiscible liquid in which the water - insoluble active ingre administration of a composition according to any of the dient is soluble ; and a hydrophilic surfactant having an HLB composition embodiments described herein . In particular , value of at least 10 , the composition being adapted to release the disorder is selected from ( i ) inflammatory bowel disease water - insoluble active ingredient in at least the colon and the and (ii ) graft -versus -host disease following hematopoietic water- insoluble active ingredient being for therapeutic use in stem cell transplantation . In particular, the disorder is colitis . combination therapy with a hydroxylase inhibitor. [ 0037 ] There is therefore provided a method for mainte 10042 ] The hydroxylase inhibitor may be comprised in a nance therapy of a subject who has suffered from or is pharmaceutical formulation adapted to release the inhibitor suffering from a disorder selected from (i ) inflammatory in at least the colon . In particular, the unit solid containing bowel disease and ( ii ) graft - versus -host disease following the water - insoluble active ingredient may contain the hematopoietic stem cell transplantation , which method com hydroxylase inhibitor; in this case the composition may prises administered to said subject a hydroxylase inhibitor. comprise a single population of beads . It is implicit , and by Particular embodiments of this method comprise the admin virtue of this sentence explicit , that the hydroxylase inhibitor istration of a composition according to any of the compo is for therapeutic use . sition embodiments described herein . [0043 ] The composition may comprise two populations of 0038 ] There is therefore provided a method for mainte unit solids, e . g . two populations of minibeads , a first popu nance therapy of a subject who has suffered from , or is lation containing the water - insoluble active ingredient and a suffering from , colitis which method comprises adminis second population containing the hydroxylase inhibitor. tered to said subject a hydroxylase inhibitor. Particular [0044 ] Further provided is an emulsion for use in manu embodiments of this method comprise the administration of facturing a composition as above , the emulsion comprising a composition according to any of the composition embodi oil droplets dispersed in an aqueous phase characterised in ments described herein . that the aqueous phase comprises a water- soluble polymer [0039 ] Aspects and embodiments of the present invention and in that the emulsion comprises a water- insoluble active relate to compositions comprising an immunosuppressant as ingredient selected from calcineurin inhibitors ,mTor inhibi an active ingredient, and more particularly water - insoluble tors, and macrolide immunosuppressants , a hydroxylase active ingredients selected from calcineurin inhibitors , m Tor inhibitor and a hydrophilic surfactant having an HLB value inhibitors and macrolide immunosuppressants . Embodi of at least 10 . ments relate to compositions comprising a macrolide immu (0045 ] A further aspect of the invention resides in a nosuppressant as active ingredient. Exemplary macrolide pharmaceutical composition for oral administration , obtain immunosuppressants cyclosporins, tacrolimus , ascomycins able by : and sirolimus , for example cyclosporin A , whose interna [0046 ] A ) mixing together at least the following materials tional non -proprietary name is ciclosporin . Macrolide to form an emulsion : immunosuppressants are poorly soluble in water but may be 10047 ) i ) a water - insoluble active ingredient selected dissolved more readily in hydrophobic environments , as from calcineurin inhibitors , mTor inhibitors, and described herein . macrolide immunosuppressants ; [0040 ] The disclosure includes an invention predicated on [0048 ] ii ) an aqueous phase comprising water and a a finding relating to minibeads which themselves form an water - soluble polymer material; embodiment of the invention and which comprise a matrix [0049 ] iii ) a hydrophobic liquid ; constituted by gelatin or another water soluble polymer and [0050 ] iv ) a hydrophilic surfactant having an HLB value contain a macrolide immunosuppressant, a water- immis of at least 10 ; cible liquid in which the macrolide immunosuppressant is [ 0051 ] V ) optionally one or more excipients which are soluble and a hydrophilic surfactant having an HLB value of miscible with or soluble in hydrophobic liquid to at least 10 . It is inferred in these beads that the water increase the solubility of the immunosuppressant in the immiscible liquid forms droplets or inclusions in thematrix liquid , ( and for the purposes of defining the invention this is wherein the water - insoluble active ingredient is soluble in considered to be the case ) , with the immunosuppressant the hydrophobic liquid when it is combined with any said mainly or wholly ( for practical purposes ) being in the one or more excipients ; and water - immiscible liquid and at least a significant proportion [0052 ] B ) formulating the emulsion into a pharmaceutical of the hydrophilic surfactant being in the water -soluble composition comprising a unit solid which comprises the polymer phase (outside the water - immiscible liquid ). Such emulsion in a dry state, wherein the composition is adapted minibeads, in some cases further containing a hydroxylase to release the active ingredient at least into the colon and the inhibitor, and having a coating designed to achieve release active ingredient is for therapeutic use in combination of the active ( s ) at least in the colon , were tested in an animal therapy with a hydroxylase inhibitor. model and found to protect against loss (resist loss ) of [ 0053 ] The invention includes a method for administering intestinal barrier function . Those tests involving co - admin to a subject ( i) a water- insoluble active ingredient selected istration of the macrolide immunosuppressant and the from calcineurin inhibitors ,mTor inhibitors and macrolide hydroxylase inhibitor were particularly effective . See in immunosuppressants ; and ( ii) a hydroxylase inhibitor , the particular Examples 1 - 5 . method comprising: [0041 ] The invention therefore includes a pharmaceutical [ 0054 ] orally administering a pharmaceutical composition composition for oral administration , comprising a unit solid comprising a unit solid which comprises a water - soluble which comprises a water -soluble polymer matrix material in polymer matrix material in which matrix material are dis which matrix material are dispersed a water - insoluble active persed the water - insoluble active ingredient , droplets of ingredient selected from calcineurin inhibitors ,mTor inhibi water - immiscible liquid in which the water - insoluble active US 2018 / 0042989 A1 Feb . 15 , 2018
ingredient is soluble and a hydrophilic surfactant having an includes the GIT below the small intestine . The small HLB value of at least 10 , the composition being adapted to intestine is the primary location of absorption of drugs and , release the water- insoluble active ingredient in at least the by preferentially delivering the active agent below the small colon ; and intestine , a therapy may reduce absorption whilst permitting [ 0055 ] simultaneous, sequentially or separately adminis effective local activity in the GIT , such as in the colon . It is tering a hydroxylase inhibitor to the subject generally understood that the GIT below the small intestine 100561. The invention also includes a method for treating a comprises the large intestine which , according to the Ter disorder of, or at least suspected of being associated with , a minologia Anatomica ( TA ) , the international standard on leaky intestinal epithelial barrier, the method comprising human anatomic terminology , comprises the cecum , colon , orally administering an effective amount of a pharmaceutical rectum and anal canal. Macrolide immunosuppressants are composition comprising a water -soluble polymer matrix poorly absorbed in the colon and targeted delivery to the material in which are dispersed a water- insoluble active colon as described herein is beneficial to treat disorders of ingredient selected from calcineurin inhibitors , mTor inhibi the colon , rectum and /or anal canal. tors and macrolide immunosuppressants, and droplets of [ 0061 ] Preferential delivery of the active agent to the GIT water - immiscible liquid in which the active ingredient is below the small intestine means that the active agent is soluble, wherein the composition is adapted to release the preferentially released into the lumen of the GIT below the active ingredient in at least the colon . See in particular small intestine , and preferably into the colon . All aspects of Examples 1 - 5 . the invention may be implemented using a formulation [0057 ] Another embodiment is a product for use in manu which comprises minibeads ( sometimes called minicap facturing a composition as previously defined , the product sules) and optionally more than half of the minibeads, e . g . being a water -soluble polymermatrix material formulated as all of them , comprise a barrier to prevent release and a minibead having a diameter of no more than 10 mm , e. g . degradation of the active agent in the stomach and small of not more than 5 mm , in which matrix material are a intestine . The barrier may comprise an enteric polymer water - insoluble active ingredient selected from calcineurin product , for example an enteric coating , or it may comprise inhibitors , m Tor inhibitors and macrolide immunosuppres an erodible coating . The barrier may comprise a coating sants ; droplets of a water - immiscible liquid in which the which comprises a polymer, e . g . a polysaccharide, which is water - insoluble active ingredient is soluble , and a hydro specifically susceptible to degradation by bacterial enzymes philic surfactant having an HLB value of at least 10 . in the colon , i. e . is susceptible to degradation by bacterial [0058 ] Further provided by the invention is a pharmaceu enzymes in the colon but not by enzymes higher up the GIT . tical composition for oral administration , comprising a [0062 ] A formulation comprising an immunosuppressant , water -soluble polymer matrix material in which matrix e .g . a macrolide immunosuppressant, may be for use in material are dispersed a water- insoluble active ingredient achieving systemic inhibition of a pro - inflammatory selected from calcineurin inhibitors , mTor inhibitors and cytokine , for example IL - 1 , IL - 17 , IL - 8 , IL - 22 and / or TNFa macrolide immunosuppressants; droplets of a water - immis which is lower than the local inhibition in the gastrointes cible liquid in which the water - insoluble active ingredient is tinal tract . The formulation may be for use in limiting soluble, wherein the composition is adapted to release the systemic inhibition of a pro - inflammatory cytokine while water- insoluble active ingredient in at least the colon and is achieving clinically effective local inhibition in the gastro for use in treating a disorder of, or at least suspected ofbeing intestinal tract, e . g . optimising the local inhibition in the associated with , a leaky intestinal epithelial barrier. In one gastrointestinal tract . See Example 5 . embodiment the disorder is not Crohn ' s disease , ulcerative 100631 For all aspects and implementations of the inven colitis , GVHD , or GI- GVHD . In another embodiment the tion , therefore , at least a portion of the active agent may be disorder is Crohn ' s disease , ulcerative colitis , GVHD , or protected against degradation in the upper gastrointestinal GI- GVHD . In one embodiment the disorder is not inflam tract , and optionally in the stomach and small intestine. The matory bowel disease , Crohn 's disease, ulcerative colitis , formulation may be adapted for a first portion of the active GVHD , or GI- GVHD . In another embodiment the disorder agent to be released in the upper gastrointestinal tract and a is inflammatory bowel disease , Crohn ' s disease , ulcerative second portion of the active agent to be released in the colon . colitis , GVHD , or GI- GVHD . In one embodiment the dis The second portion may comprise more than half said active order is not irritable bowel syndrome, inflammatory bowel agent in the formulation , e . g . at least 60 % , at least 70 % or disease , Crohn ' s disease , ulcerative colitis , GVHD , or GI at least 80 % thereof . As mentioned above , therefore , the GVHD . In another embodiment the disorder is irritable formulation may comprise minibeads and more than half of bowel syndrome, inflammatory bowel disease , Crohn ' s dis the minibeads, e . g . at least 60 % of them , at least 70 % of ease , ulcerative colitis , GVHD , or GI- GVHD . Also dis them , at least 80 % of them or all of them , may comprise a closed is an embodiment in which the disorder is not irritable barrier to prevent release and degradation of the active agent bowel syndrome, inflammatory bowel disease, Crohn 's dis in the stomach and small intestine. The protection against ease , ulcerative colitis , GVHD , or GI- GVHD . degradation or barrier may be provided by a coating selected [ 0059 Macrolide immunosuppressants are water- in from enteric coatings and coatings comprising a polymer soluble and act locally in the colon to suppress inflamma specifically susceptible to degradation by bacterial enzymes tions, and non -macrolide ( as well as macrolide ) calcineurin in the colon . inhibitors and mTor inhibitors have the same function and 10064 ] However, celiac disease may also manifest itself in are therefore usable in place of a macrolide immunosup the colon as microscopic colitis and IBD . Further , patients pressant . suffering from celiac disease suffer an increased risk of also [0060 ] For all aspects and implementations of the inven suffering from ulcerative colitis . It will therefore often be tion , the therapy may affect the entirety of the GIT or a beneficial for celiac patients to have the actives described portion thereof, in particular any portion thereof which herein delivered to the colon as well as the small intestine . US 2018 / 0042989 A1 Feb . 15 , 2018
The invention therefore provides combination formulations the minibeads, i . e . the minibeads may have the characteris comprising hydralazine and cyclosporin A which are for use tics ofminibeads during the process of manufacturing which in delivery of the actives in the small intestine and for use the hydralazine was dissolved in an aqueous phase mix or in combination with a combination formulation comprising premix to form a clear solution , and the minibeads may be hydralazine and cyclosporin A for us in delivery of the obtained by such a process. The minibeads may include a actives in the colon . The two different combination formu dispersed phase , e . g . to include a hydrophobic active , but a lations may be administered simultaneously, sequentially or dispersed phase is not necessary , and the minibeads may separately . For example , a unit dosage , e . g . a capsule , may therefore be optionally coated single -phase (matrix -only ) comprise a first population of hydralazine/ cyclosporin mini minibeads. This insight applies both to hydralazine for capsules having a controlled release coating to release the maintenance therapy and therefore optionally for use in active in the small intestine and a second population of monotherapy, and to hydralazine for combination therapy as hydralazine / cyclosporin minicapsules having a controlled described herein , e . g . with an immunosuppressant, to restore release coating to release the actives in the colon . See below or improve GIT health . Any immunosuppressant may be for more about coatings . The second population may con included in the same minibeads as the hydralazine , in a stitute no more than 50 % of minibeads in the unit dosage second population of minibeads from the hydralazine in the form , e. g . at least 10 % or at least 20 % and no more than 30 % same dosage form , or in a separate dosage form , e . g . in or 40 % , for example from 20 % - 40 % or 10 % -30 % . separate multiple minibead capsules . Specifically a unit [0065 ] The active agent, therefore, may all be comprised dosage form , for example a multiple minibead capsule may in minibeads. A multiplicity of such minibeads may be comprise hydralazine in an amount of from 2 . 5 mg- 100 mg, comprised in a unit dosage form , for example a gelatine or e . g. 2 .5 mg- 50 mg or 5 mg- 100 mg , e .g . 5 mg- 50 mg. The other capsule, a sachet or a compressed tablet. The formu amount of hydralazine may be from 2 .5 mg to 25 mg, e. g . lation may therefore consist of a multiplicity of minibeads 5 mg- 25 mg, for example 5 mg- 10 mg. The unit dosage form and optionally a capsule or other container for the mini may be administered for example one , two or three times a beads. For all aspects and implementations of the invention , day, e . g . may be for once a day therapy . In particular , the unit the formulation may comprise a first population of mini dosage forms described in this paragraph are for adminis beads adapted to release the active agent in the upper tration to human subjects . gastrointestinal tract and a second population of minibeads 10068 ] One embodiment of the minibeads mentioned adapted to release the active agent in the colon . The second herein comprises a water -soluble polymer matrix in which population of minibeads may comprise more than half said are dispersed droplets of water - immiscible liquid , the matrix active agent in the formulation , e. g . at least 60 % , at least including a surfactant and the composition comprising a said 70 % or at least 80 % thereof. The second population of active principle . In another embodiment, the minibeads minibeads may have a coating comprising a polymer spe comprise a water - soluble polymer matrix in which are cifically susceptible to degradation by bacterial enzymes in dispersed droplets of water - immiscible liquid , the water the colon , i .e . susceptible to degradation by bacterial immiscible liquid comprising a surfactant and the compo enzymes in the colon but not by enzymes higher up the GIT. sition comprising a said active principle . In a further [0066 ] The formulation may comprise multiple seamless embodiment, the minibeads comprise a water -soluble poly minibeads ( also known as seamless minicapsules) compris mer matrix in which are dispersed droplets of water- immis ing the active agent. The minibeads referred to throughout cible liquid , the matrix including a surfactant, the water this specification may therefore be seamless . The seamless immiscible liquid comprising a surfactant, and the minibeads may comprise a water soluble polymer matrix composition comprising a said active principle . and , dispersed in the matrix , a dispersed phase composed of [0069 ] The dispersed phase described herein may be an oil materials selected from hydrophobic and amphiphilic mate phase. The dispersed phase described herein may be a rials , and combinations thereof . The minibeads may com micellar phase . prise a composition having the characteristics of a dried state [ 0070 ] The extent to which dissolution may affect the of colloid having a continuous aqueous phase comprising a composition ' s physical form and features depends on the hydrogel- forming polymer . The colloid may have a dis initial shape , size and make -up of the composition . Where persed phase selected from a hydrophobic phase , a water in the composition bears a coat , the rate and manner of oil emulsion , and a micellar phase selected from micelles, dissolution can be modified ( see below ) . promicelles and combinations thereof. The matrix , or the 10071 ] In one aspect, the minibeads can be described as hydrogel- forming polymer of the dried colloid , may com comprising a dried oil- in - water ( o / w ) emulsion . The formu prise a hydrophilic surfactant. The dispersed phase may lations of the invention may comprise multiple droplets of comprise a hydrophobic surfactant. For all aspects and water- immiscible liquid within a moulded or shaped form implementations of the invention , the matrix ( the hydrogel e . g . a minibead . The formulations of the invention may forming polymer) may comprise a hydrophilic surfactant comprise multiple droplets of surfactant within a moulded or having an HLB value of at least 15 and the dispersed phase shaped form e . g . a minibead . may be a hydrophobic phase comprising a non - ionic sur [ 0072 ] The formulations of the invention may comprise a factant having an HLB value of at least 10 but less than that plurality of optionally coated minibeads having a water of the hydrophilic surfactant. soluble polymer matrix . In a particular embodiment, the [0067 ] It is believed having regard to the data included in formulation comprises a plurality of minibeads of dried this application that a multiple minibead formulation as oil - in -water emulsion . described herein and wherein the minibeads comprise [0073 ] In the case of formulations which comprise mini hydralazine , may comprise a low amount of hydroxylase to beads, at least some of the minibeads ( e . g . a first population ) be clinically useful. In particular, the hydralazine in such a may comprise a said active agent (optionally in combination formulation may be in solid solution in the matrix phase of with at least one further active ingredient) and optionally US 2018 / 0042989 A1 Feb . 15 , 2018 other minibeads ( e . g . a second population ) which comprise solid or solidified external phase . The solid external phase a said active agent (optionally in combination with at least dissolves on contact with an aqueous medium . one further active ingredient) or one population may be free 10080 ] The term “ matrix ” is a term well -known in the art of active principles or include “ deactivating ” principles e . g . and generally means , according to context, a solid , semi enzyme or toxin sequesters or include active excipients , solid , undissolved or not - yet - dissolved material which pro such as, for example, permeability enhancers, which may vides structure and volume to a composition . enhance , moderate or potentiate the effect of an active 10081 ] Solidification of the external phase may have arisen principle in another population . The formulations of the through various means including chemically ( e . g . by cross invention may comprise multiple populations ofminibeads . linking ) or physically ( e . g . by cooling or heating ) . By use of The active principles may be the same or different as the term “ dried ” , it is not sought to imply that a drying step between populations, provided that at least one population is necessary to produce the dried emulsion or micelle includes an active agent as required by the invention . composition ( although this is not excluded ) rather that the [0074 ] The invention includes formulations in which an solid or solidified aqueous external phase is substantially active agent and optionally one or more further active free of water or free of available water. In this respect , the ingredient( s ) is ( are ) incorporated in a dispersed phase term " aqueous phase ” is nevertheless employed in this mentioned herein . The invention includes formulations in document to denote the external ( continuous ) phase of the which an active agent and optionally one or more further composition of the invention even though water , in certain active ingredient ( s ) is (are ) incorporated in a continuous embodiments , is largely absent from (or trapped within the phase (matrix phase ) mentioned herein . Both the dispersed cross - linked matrix of) the minibead compositions. The phase and the continuous phase may include an active external phase of the minibead composition is however ingredient, and each phase may have an active ingredient water- soluble and dissolves in aqueous media . In one different from that in the other phase, the minibeads com embodiment , the oil droplets or micelles are released when prising an active agent required by the invention . the aqueous phase dissolves or is exposed to aqueous media . [ 0075 ]. In the case of formulations comprising minibeads, [0082 ] The term “ released ” in relation to the oil droplets or the minibeads may be coated with a polymer to alter the micelles means free to move , egress , coalesce , dissolve , release profile or to protect the bead and/ or the active ( re )emulsify etc . although actual movement, egression , principle within the bead from degradation or oxidation or coalescence , association or ( re ) emulsification is not a hydrolysis or proteolysis or degradation mediated by high or requirement i . e . may not occur and indeed may intentionally low pH . be constrained e . g . by presence of a coat or coating and / or [0076 ] In the instance of formulations comprising mini by incorporation of certain constraining or retarding sub beads having a water - soluble polymer matrix and a dis stances into the water - soluble polymer matrix . persed phase comprising a hydrophobic material , an amphi [0083 ] The inclusion in the aqueous phase of a surfactant philic material or a combination thereof, the invention is of ( described below ) may lead to improved dissolution or particular interest for active agents of low aqueous solubility release of the active agent. In particular, it has been found and / or liposoluble active agents where incorporation into the that, when the formulation comprises minibeads bearing a dispersed phase ( e . g . an oil phase ) brings particular advan polymeric coating , inclusion of a surfactant in the aqueous tages . phase enhances dispersion / egress through pores or other [0077 ] The invention includes within its scope formula openings in the polymer coat ( or other local removal, tions in which a said active agent is formulated for oral swelling or weakening of the polymer coat ). Where the administration as minibeads of dried oil - in -water emulsions minibeads have an oil phase which comprises a surfactant, in which the active agent may be incorporated in the oil the surfactant included in the aqueous phase may be different phase of the emulsion , the beads being optionally coated from any surfactant included in the oil phase . with a polymer . The invention includes within its scope [0084 ] In embodiments , the formulations comprise mini formulations in which a said active agent is formulated for beads which comprise a matrix constituted by gelatin or oral administration as minibeads of dried aqueous micelle another water soluble polymer (or a combination thereof) compositions in which the active agent may be incorporated and contain a water -insoluble active agent, a water- immis in the micelle phase of the emulsion , the beads being cible liquid in which the active agent is soluble and a optionally coated with a polymer. hydrophilic surfactant having an HLB value of at least 10 . [0078 ] The water- soluble polymer matrix ( or in one It is inferred in these minibeads that the water - immiscible aspect, the aqueous phase of a dried emulsion ) comprises , in liquid forms droplets or inclusions in the matrix ( and for the one embodiment, a cross - linked water- soluble polymer e . g . purposes of defining the invention this is considered to be resulting from chemical or physico -chemical (e . g . drying ) the case ) , with the active agent mainly or wholly ( for solidification of a fluid aqueous continuous phase such that, practical purposes) being in the water - immiscible liquid and in the matrix or dried emulsion , water is substantially absent at least a significant proportion of the hydrophilic surfactant and the dispersed phase is immobilized . In this embodiment, being in the water - soluble polymer phase ( outside the water the dried aqueous phase can therefore be referred to as an immiscible liquid ). immobilization matrix . [0085 ] Some formulations of the invention are obtainable [ 0079 ] The term “ dried emulsion ” generally means an by a process comprising : emulsion whose internal ( discontinuous ) phase has been [0086 ] B ) mixing together at least the following materials immobilized in a substantially solid or solidified external to form an emulsion : phase . The solid external phase dissolves on contact with an [ 0087 ] i) a said active agent; aqueous medium . The term “ dried aqueous micelle compo [0088 ] ii ) an aqueous phase comprising water and a sition ” generally means an aqueous micelle composition water - soluble polymer material ; whose micelle phase has been immobilized in a substantially [ 0089 ] iii ) a hydrophobic liquid ; US 2018 / 0042989 A1 Feb . 15 , 2018
[0090 ] iv ) a hydrophilic surfactant having an HLB value ture . It is inferred therefore in such embodiments that , in the of at least 10 ; final minibead , the water - insoluble active ingredient is in [ 0091 ] v ) optionally one or more excipients which are solution in the water immiscible liquid . Similarly , if a miscible with or soluble in hydrophobic liquid to water - soluble active ingredient is used , it will in the embodi increase the solubility of the immunosuppressant in the ments mentioned in this paragraph be dissolved in the liquid , aqueous phase and it is inferred that the water soluble active wherein the water- insoluble active ingredient is soluble in ingredientmay in this case be considered in the minibead to the hydrophobic liquid when it is combined with any said be in solid solution in the polymer ( i. e . outside the dispersed one or more excipients , and phase ) . [ 0092 ] C ) formulating the emulsion into a pharmaceutical [0109 ] The invention includes embodiments in which composition comprising a minibead which comprises the water and water - soluble constituents are formed into a clear emulsion in a dry state , wherein the composition is adapted solution which is mixed with the dispersed phase medium to release the active ingredient at least into the colon . ( e . g . a water immiscible liquid ) . The invention includes [0093 ] For all embodiments and aspects of the invention , embodiments in which a water- immiscible liquid and water the minibead may optionally have the characteristics of a insoluble constituents are formed into a clear solution which product prepared by a method comprising : is mixed with the aqueous phase . Both phases may be clear [0094 ] A ) mixing together at least the following materials solutions prior to mixing of them to form an emulsion . to form an emulsion : [0110 ] The reader will understand that the manufacture [0095 ] i ) the active agent( s ) ; may use water and a water - immiscible liquid , e . g . a liquid [0096 ] ii ) an aqueous phase comprising water and a lipid , or water and a micelle - former ( surfactant ) . During water - soluble polymer material ; manufacture , the constituents will partition between the [ 0097 ) iii ) a hydrophobic liquid ; water phase and the non - aqueous phase , or their interface , [0098 ] iv ) a hydrophilic surfactant having an HLB value according to their solubilities in each phase . The water of at least 10 ; insoluble constituents will overwhelmingly be present in the 00991 v ) optionally one or more excipients which are water- immiscible part (water immiscible liquid or micelle miscible with or soluble in hydrophobic liquid to interior) and water- soluble but lipid - insoluble constituents increase the solubility of the immunosuppressant in the will overwhelmingly be present in the water phase . Absolute liquid , exclusion of a constituent from one phase down to the [ 0100 ] wherein the water - insoluble active ingredient is molecular level may not occur. Some constituents , e . g . soluble in the hydrophobic liquid when it is combined with surfactants , may be significantly soluble in both phases and any said one or more excipients ; and may be significantly present in both phases , though they may [ 0101 ] B ) formulating the emulsion into a unit solid which predominantly be in one of the two phases . Thus , where it comprises the emulsion in a dry state . is stated that a hydrophilic surfactant is in the aqueous phase , [ 0102] In particular the mixing together may optionally it may be expected that a small portion of the surfactant may comprise forming a clear solution of hydrophobic (water perhaps be found in the non -aqueous phase . The same insoluble ) active agent( s ) in the hydrophobic liquid together applies mutatis mutandis to hydrophobic surfactants . with any said one or more excipients. For all aspects and [0111 ] It is inferred that when the liquid emulsion has been implementations of the disclosure , there may be formed a converted to a solid , e . g . a minibead , the various constituents premix of the continuous phase and a premix formed of the will remain partitioned between the non - aqueous phase and disperse phase , each complete premix optionally being a the dried aqueous phase ( i. e . the water - soluble polymer clear solution before the two premixes are combined to form matrix ) substantially in the samemanner as in the emulsion . a liquid colloidal composition , e . g . emulsion or aqueous This has not been demonstrated , however . micelle composition . [0112 ] The compositions of the invention may be for use [0103 ] More particularly , the manufacture of a minibead in treating a disorder of, or at least suspected of being may optionally comprise : associated with , a leaky intestinal epithelial barrier . [0104 ] i ) forming an aqueous phase premix comprising , [0113 ] This paragraph applies to all aspects and imple or usually consisting of, a solution in water of water mentations of the disclosure . In one embodiment the disor soluble constituents including a water -soluble thermo der is not Crohn ' s disease , ulcerative colitis , GVHD , or tropic hydrogel- forming polymer, any water- soluble GI- GVHD . In another embodiment the disorder is Crohn ' s excipient( s ), any water- soluble active ( s ) ; disease , ulcerative colitis , GVHD , or GI- GVHD . In one [0105 ] ii ) forming a water - immiscible phase premix embodiment the disorder is not inflammatory bowel disease , ( sometimes called an oil phase premix ) comprising , or Crohn ' s disease , ulcerative colitis , GVHD , or GI-GVHD . In usually consisting of, a solution in water - immiscible another embodiment the disorder is inflammatory bowel liquid of water - insoluble constituents ( e . g . disease , Crohn ' s disease , ulcerative colitis , GVHD , or GI water - insoluble active ( s ) ) ; GVHD . In one embodiment the disorder is not irritable f0106 ] iii ) mixing the two phases form an emulsion , and bowel syndrome, inflammatory bowel disease , Crohn ' s dis 10107 ] iv ) formulating the emulsion into a minibead , ease, ulcerative colitis , GVHD , or GI-GVHD . In another e . g . ejecting it through a single orifice nozzle to form embodiment the disorder is irritable bowel syndrome, droplets which are caused or allowed to fall into a inflammatory bowel disease , Crohn ' s disease, ulcerative water- immiscible cooling liquid in which the droplets colitis , GVHD , or GI- GVHD . cool to form minibeads, and then separating the mini [0114 ] A judicious combination of different types of poly beads from the cooling liquid . meric coating (described in more detail below ) can produce 10108 ] In embodiments, therefore, the one or more active advantages in relation to the in vitro dissolution and in vivo pharmaceutical ingredients are dissolved during manufac performance of the composition of the invention . In particu US 2018 / 0042989 A1 Feb . 15 , 2018 lar, inclusion of a polymer which degrades in the presence [0125 ] FIG . 7 is weight loss change graph relating to of bacterial enzymes present in the colon ( and /or a polymer hydralazine administered orally or i. p . ( see the comparative which encourages the formation of pores in the coating - a example ) ; " pore - former” ) with a pH - independent polymer leads to [0126 ] FIG . 8 is a Disease Activity Index (DAI ) graph release of active principle substantially in the colon or other relating to hydralazine administered orally or i. p . ( see the pre -determined site of the GI tract . In a particular embodi comparative example ) ; ment, the abovementioned polymer degradable by bacterial 0127 ] FIG . 9 is a graph of Transepithelial Electrical enzymes is water- soluble . Resistance ( TEER ) showing the effects of hydralazine at [0115 ] Throughout the description and claims of this different concentrations in aqueous solutions with DSS specification , the words “ comprise ” and “ contain " and varia ( dextran sodium sulphate ) ( see Example 4 ) ; tions of them mean “ including but not limited to ” , and they [0128 ] FIG . 10 comprises a photograph of a gel illustrat are not intended to ( and do not) exclude other moieties , ing changes in HIF - la expression using hydralazine in additives , components, integers or steps. Throughout the Caco - 2 cells (see Example 4 ) ; description and claims of this specification , the singular 101291. FIG . 11 is a bar chart showing the number of encompasses the plural unless the context otherwise adenocarcinomas in colons of IL - 10 - - mice administered requires . In particular, where the indefinite article is used , various compositions and wherein " SmPill” designates a the specification is to be understood as contemplating plu formulation of the invention and “ CyA ” means Cyclosporin rality as well as singularity , unless the context requires A ( also known as ciclosporin ) ; otherwise . [0116 ] Features, integers , characteristics, compounds , [0130 ] FIG . 12 . is a graph showing the change in weight chemical moieties or groups described in conjunction with a of IL - 10 - ) - mice with different treatment regimes over 6 particular aspect , embodiment or example of the invention weeks (see Example 5 ) ; are to be understood to be applicable to any other aspect, [0131 ] FIG . 13 is a bar chart showing the weight change embodiment or example described herein unless incompat of IL - 10 - / - mice with different treatment regimes on day of ible therewith . All of the features disclosed in this specifi termination (day 42 ) as percentage weight change relative to cation ( including any accompanying claims, abstract and starting weight ( at week 0 of experiment ) ( see Example 5 ) ; drawings ) , and /or all of the steps of any method or process [0132 ] FIG . 14 is a bar chart showing the Disease Activity so disclosed , may be combined in any combination , except Index (DAI ) in IL - 10 - / - mice with different treatment combinations where at least some of such features and / or regimes on day of termination ( see Example 5 ) ; steps are mutually exclusive. Accordingly every disclosure 0133 ] FIG . 15 is a bar chart showing the level of Serum herein is applicable to every aspect and implementation of Amyloid A (SAA ) in IL - 10 - - mice with different treatment the invention , unless mutually exclusive . The invention is regimes at the day of termination ( see Example 5 ) ; not restricted to the details of any foregoing embodiments . [0134 ] FIG . 16 is a bar chart showing the numbers of The invention extends to any novel one , or any novel adenocarcinomas in colons of IL - 10 - / - mice with different combination , of the features disclosed in this specification treatment regimes at the day of termination ( see Example 5 ); ( including any accompanying claims, abstract and draw [0135 ] FIG . 17 is a bar chart showing myeloperoxidase ings ), or to any novel one, or any novel combination , of the (MPO ) enzymatic activity in the colons of IL - 10 - ) - mice steps of any method or process so disclosed . with different treatment regimes (see Example 5 ) ; [ 0117 ] The reader ' s attention is directed to all papers and [0136 ] FIGS. 18A , 18B and 18C are bar charts showing documents which are filed concurrently with or previous to the colon levels of cytokines TNF - a and IL - 17 in IL - 10 - 1 this specification in connection with this application and mice with different treatment regimes (see Example 5 ) ; which are open to public inspection with this specification , [0137 ] FIG . 19 is a bar chart showing histology scores of and the contents of all such papers and documents are proximal colons of IL - 10 - - mice with different treatment incorporated herein by reference . regimes ( see Example 5 ) ; 10138 ] FIGS. 20A , 20B and 20C are bar charts showing BRIEF DESCRIPTION OF THE DRAWINGS the in vitro production of cytokines TNF - a and IL - 17 by 0118 ] Embodiments of the invention are further described spleen cells from IL - 10 - - mice with different treatment hereinafter with reference to the accompanying drawings , in regimes (see Example 5 ) ; which : 101391. FIG . 21 is a graph showing % weight change [ 0119] FIG . 1 is a plot of weight loss change showing the resulting from different treatment regimes in a DSS model of effect of compositions of the invention ( see Example 2 ) ; intestinal fibrosis in mice ( see Example 6 ); [0120 ] FIG . 2 is a Disease Activity Index (DAI ) graph [0140 ] FIG . 22 is a graph showing change of DAI result showing the effect of compositions of the invention ( see ing from different treatment regimes in the DSS model of Example 2 ) ; intestinal fibrosis in mice ( see Example 6 ) ; [0121 ] FIG . 3 is a set of histology slides showing the effect 10141 ] FIG . 23 is a bar chart showing the ratio of colon of compositions of the invention (see Example 2 ); weight: colon length resulting from different treatment [0122 ] FIG . 4 is a second plot of weight loss change regimes in the DSS model of intestinal fibrosis in mice ( see showing the effect of compositions of the invention ( see Example 6 ) ; Example 3 ); [0142 ] FIG . 24 is a graph showing % weight change [0123 ] FIG . 5 is a second Disease Activity Index (DAI ) resulting from treatment with formulations containing dif graph showing the effect of compositions of the invention ferent proportions of hydralazine and ciclosporin in a DSS ( see Example 3 ) ; model of colitis in mice and wherein “ Hya” means hydrala [ 0124 ] FIG . 6 is a second set of histology slides showing zine and " Cya ” means cyclosporin A ( also known as the effect of compositions of the invention (see Example 3 ); ciclosporin ) ( see Example 7 ) ; US 2018 / 0042989 A1 Feb . 15 , 2018
[0143 ] FIG . 25 is a graph showing change of DAI result - a maximal activation at 10 ug /mL and lower signals by ing from treatment with formulation containing different increasing the concentration of the drug . An explanation is proportions ofhydralazine and ciclosporin in the DSS model that the concentration of drug higher than 100 ug /mL could of colitis in mice and wherein “ Hya ” means hydralazine and kill the cells . The activation range of NFKB seems to be " Cya ” means cyclosporin A ( see Example 7 ) ; between 0 and 10 ug /mL of drug . At all concentrations the [0144 ] FIG . 26 is a bar chart showing the ratio of colon excipients do not have any effect on the cells survival. weight: colon length resulting from treatment with formula [0160 ] In FIG . 30 no significant activation of NFKB tion containing different proportions of hydralazine and related to the drug is shown . Both forms result to give a ciclosporin in the DSS model of colitis in mice and wherein similar activation level of NFKB , demonstrating again the “ Hya ” means hydralazine and " Cya ” means cyclosporin A stability of the formulated drug . (see Example 7 ) . [0161 ] In FIG . 31 both drugs are administrated together at [ 0145 ] FIG . 27 shows the effects on LPS - induced activa - the same concentrations. The greater influence of hydrala tions of NFKB by cyclosporin A and hydralazine ( drug A ) ; zine HCl on the NFKB activation is clearly recognizable . To 10146 ) FIG . 28 shows the effects on LPS - induced activa note is that the signal of formulated drug is greater than tion of IL - 8 by cyclosporin A , hydralazine ( drug A ) ; those of unformulated drug ( except for 10 ug /mL and 25 [0147 ] FIG . 29 is a bar chart showing normalised values of ug /mL ) . This could indicate a better stability of the com NFKB activation attained by formulated and unformulated bined drugs in the formulated form , thanks the delivery in hydralazine as measured using an NRE - luciferase assay ; different phases . In the formulation Cyclosporin A is dis [0148 ] FIG . 30 is a bar chart showing normalised values of solved in the oil phase and cannot interact with hydralazine , NFKB activation attained by formulated and unformulated dissolved in the water phase . The interaction of the two cyclosporin A as measured using an NRE - luciferase assay ; drugs is instead possible in the unformulated form . The [0149 ] FIG . 31 is a bar chart showing normalised values of concentration values in FIG . 31 express that of hydralazine NFKB activation attained by formulated and unformulated ( in the combination formulation ) . combination of hydralazine and cyclosporin A ; [0150 ] FIG . 32 comprises histology slides showing the DETAILED DESCRIPTION effect of different treatment regimes in the DSS model of [0162 ] The present methods and compositions are as pre intestinal fibrosis in mice to which FIGS. 21 -23 relate (see viously described useful for treatment of disorders of the GI Example 6 ) ; tract which involve or result at least in part from inflamma [0151 ] FIG . 33 comprises slides showing the histology of tory conditions, the treatments including by way of example healthy control specimens of colonic tissue in the DSS maintenance therapy or prophylaxis as well as treatment to model of colitis in mice to which FIGS . 24 - 26 relate ( see improve the condition of a patient . The methods and com Example 7 ) ; positions are useful for treating such GI tract conditions [ 0152 ] FIG . 34 comprises slides showing the histology of whilst limiting absorption of the active ( s ) and thus limiting DSS control specimens of colonic tissue in the DSS model also unwanted system activity of the active ( s ) . Asmentioned of colitis in mice to which FIGS. 24 - 26 relate ( see Example in more detail elsewhere, methods and compositions of the 7) ; invention may relate to fibrotic disorders of the GI tract; the 10153 ] FIG . 35 comprises slides showing the histology of aspects and implementations of the invention applicable to specimens of colonic tissue from mice treated with " low fibrotic disorders are therefore equally applicable to gastro level ” hydralazine/ cyclosporin A combination minibeads in intestinal fibrosis . the DSS model of colitis in mice to which FIGS. 24 - 26 relate [0163 ] The invention will now be described in detail by (see Example 7 ) ; reference to the various components which the composition [ 0154 ] FIG . 36 comprises slides showing the histology of of the invention may comprise . The term " excipient” may be specimens of colonic tissue from mice treated with “ high used occasionally to describe all or some of the components level ” hydralazine/ cyclosporin A combination minibeads in other than the active principle (s ) bearing in mind that some the DSS model of colitis in mice to which FIGS . 24 - 26 relate excipients can be active and that some active principles can ( see Example 7 ) ; have excipient character. [0155 ] FIG . 37 is a photograph of a nitrocellulose mem [0164 ] If not otherwise stated , ingredients , components, brane stained to show HIF -1a from a lysate of Caco2 cells excipients etc of the composition of the invention are incubated with hydralazine HCl (see Example 9 ) ; suitable for one or more of the intended purposes discussed [0156 ] FIG . 38 is a photograph of a nitrocellulose mem elsewhere herein . brane stained to show HIF - la from a lysate of Caco2 cells [0165 ] For the avoidance of doubt, it is hereby stated that incubated with cyclosporin A ( see Example 9 ) ; the information disclosed earlier in this specification under [0157 ] FIG . 39 is a photograph of a nitrocellulose mem the heading “ Background ” is relevant to the invention and is brane stained to show HIF - la from a lysate of Caco2 cells to be read as part of the disclosure of the invention . incubated with a combination of hydralazine HCl and [0166 ] The treatments provided by this invention may cyclosporin A (see Example 9 ). include any one or more of maintaining the health of the GIT [0158 ] FIG . 11 shows that there were differences between ( gastrointestinal tract) , restoring or improving the health of groups in the mean number of adenocarcinomas detected in the GIT and delaying the progression of a disorder of the the colon , with CyA ( ip ) and SmPill (Cya ) groups having GIT . In particular, the invention concerns inflammatory relatively fewer. health of the GIT and the treatment of inflammatory disor [0159 ] In FIG . 29 both hydralazine HCl forms result to ders of the GIT . The term " treatment” , and the therapies give a similar activation level of NFKB with a tendency for encompassed by this invention , include the following and stronger activation by the unformulated form , demonstrating combinations thereof: ( 1 ) inhibiting , e . g . delaying initiation the stability of the formulated drug . The general trend shows and /or progression of a disorder or condition ; (2 ) preventing US 2018 / 0042989 A1 Feb . 15 , 2018 or delaying the appearance of clinical symptoms of a state , useful for patients who have suffered from , do suffer from or disorder or condition developing in an animal ( e . g . human ) have risk factors for, such cancers . The disclosed therapies that may be afflicted with or predisposed to the state , and products may be used in ( e . g . as part of ) the treatment disorder or condition but does not yet experience or display of such carcinomas or in maintenance therapies of patients clinical or subclinical symptoms of the state , disorder or who have suffered from such carcinomas . The invention in condition ; ( 3 ) inhibiting the state , disorder or condition ( e . g . , its aspects and implementations is applicable to the disorders arresting , reducing or delaying the development of the mentioned in this paragraph . disease , or a relapse thereof in case of maintenance treat 0171 ] Solubilities of compounds, e . g . actives , in a solvent ment, of at least one clinical or subclinical symptom ( for example water )may be defined as follows, the solubility thereof) ; and / or ( 4 ) relieving the condition ( i. e ., causing being measured at 25° C . and parts being by weight: regression of the state , disorder or condition or at least one of its clinical or subclinical symptoms) . The benefit to a patient to be treated may be either statistically significant or Descriptive Team Parts of Solvent for 1 part of solute at least perceptible to the patient or to the physician . It will Very Soluble Less than 1 be understood that a medicament will not necessarily pro Freely Soluble From 1 to 10 duce a clinical effect in every patient to whom it is admin Soluble From 10 to 30 istered , and this paragraph is to be understood accordingly . Sparingly Soluble From 30 to 100 Slightly Soluble From 100 to 1000 The compositions and methods described herein are of use Very Slightly Soluble From 1000 to 10 , 000 for therapy and/ or prophylaxis of the mentioned conditions. Practically Insoluble More than 10 ,000 [ 0167 ] The treatments may include maintenance therapy of patients who have suffered a GI tract disorder and whose [0172 ] Typically , but not necessarily , the invention pro condition has subsequently improved , e . g . because of treat vides that active agents which are practically insoluble , very ment. Such patients may or may not suffer a symptomatic slightly soluble or sparingly soluble in water are in the form GIT disorder. Maintenance therapy aims to arrest, reduce or of a liquid , semi- solid or solid solution in a hydrophobic or delay ( re -) occurrence or progression of a GIT disorder. amphiphilic environment, e . g . medium . [0168 ] The invention primarily concerns the treatment of [0173 ] Actives which are not particularly water soluble , humans but other warm -blooded animals , e. g. mammals are e . g . are practically insoluble , very slightly soluble or slightly also embraced by the invention , for example agricultural soluble , perhaps even are sparingly soluble , may be more mammals and domesticated mammals . Examples are pigs , soluble in a suitable dispersed phase of a minibead than in dogs and cats . For example , the compositions and methods the aqueous phase , and may therefore advantageously be of the invention may be applied to porcine proliferative incorporated in the dispersed phase . enteropathy . 0174 ] The invention provides amongst other things oral 0169 ] In some embodiments , the subject is suffering from pharmaceutical formulations for use in a therapy as men Crohn ' s disease . In other embodiments, the subject is suf tioned herein . The formulations comprise an active agent as fering from ulcerative colitis . In still further embodiments , specified previously . The formulations may be multiple the subject is suffering from irritable bowel syndrome ( e . g . minibead formulations, i . e . comprise a multiplicity of mini with constipation , diarrhea and / or pain symptoms) , celiac beads, for example at least 25 minibeads , e . g . at least 50 disease , stomach ulcers , diverticulitis , pouchitis , proctitis , minibeads . mucositis , radiation -associated enteritis , short bowel dis 10175 ] In particular embodiments , the formulations com ease , or chronic diarrhea . The administration of a composi prise minibeads and the minibeads comprise or consist of tion of the disclosure may reduce the symptoms of disease minibeads in which the content of said active agent ( s ) is in ( e . g . reduces the symptoms of inflammatory bowel disease , dissolved form , i .e . is in solution . Crohn ' s disease , ulcerative colitis, irritable bowel syndrome, 0176 ] The formulations may comprise a water -soluble celiac disease , stomach ulcers , diverticulitis , pouchitis , proc polymeric matrix in which said active agent ( s ) is or are titis , or chronic diarrhea ) . The subject may be suffering from dispersed , the matrix in particular forming minibeads which GVHD . As used herein , “ GVHD ” in particular means GI may additionally comprise one or more coatings . The poly GVHD ( gastrointestinal graft - versus -host disease ) . Further mer material constituting the matrix may be , or may com examples of inflammatory disorders to which the invention prise , a hydrogel- forming polymer . The polymer part of the may be applied are : diversion colitis , ischemic colitis , infec matrix may therefore consist of a hydrogel - forming poly tious colitis , chemical colitis , microscopic colitis (including mer. The formulations may comprise minibeads formed of a collagenous colitis and lymphocytic colitis ) , atypical colitis , polymeric matrix phase and a dispersed phase, although a pseudomembraneous colitis , fulminant colitis, autistic dispersed phase may optionally be dispensed with , in par enterocolitis , interdeminate colitis , Behcet ' s disease , ticular where the active agent ( s ) is / are water soluble such jejunoiletis, ileitis , ileocolitis and granulomatous colitis . The that they can be dissolved in the matrix phase . Alternatively , invention in its aspects and implementations is applicable to water soluble actives may be dispersed in the polymeric the disorders mentioned in this paragraph , therefore . matrix in particulate form , e . g . as micro - or nano -particles . [0170 ] The products , methods and uses of the invention The term " water soluble ” in this paragraph and elsewhere in may be applied to enteropathies , for example gluten - sensi the specification includes reference to substances which are tive enteropathy , hemorrhagic enteropathy, protein - losing categorised as soluble , freely soluble and very soluble . It enteropathy, radiation enteropathy , HIV -enteropathy , may include reference to substances where are sparingly enteropathy associated with T - cell lymphoma, autoimmune soluble . enteropathy or porcine proliferative enteropathy . Colorectal (0177 The matrix may include in addition to the water carcinoma and adenocarcinoma are inflammation - related soluble polymer and any dissolved active ( s ) , other ingredi diseases. The treatments and products described herein are ents such as, for example , excipients which may , for US 2018 / 0042989 A1 Feb . 15 , 2018 13 example ,modulate the behaviour of the matrix phase and /or a non - ionic surfactant having an HLB value of at least 10 but of other constituents during manufacture and /or after admin - less than that of the hydrophilic surfactant. The non - ionic istration . surfactant may comprise a poly ( oxyethylene) group , e . g . [ 0178 ] The matrix advantageously comprises a hydro comprise a glycerol polyethylene glycol ricinoleate ( as in philic surfactant having an HLB value of at least 10 and the case of Cremophor EL ) . particularly of at least 15 . [0186 ] In embodiments , the hydrophilic surfactant is [ 0179 ] In embodiments , the active agent does not consist selected from cationic and non - ionic surfactants , and com of a single one of, or consist of a combination consisting of, binations thereof. cyclosporin , tacrolimus , sirolimus , azathioprine , 6 -mercap [0187 ] The dispersed phase may be composed of, or topurine , methotrexate , soluble TNF receptor, antibodies predominantly of, hydrophobic and / or amphiphilic materials raised to TNF, zinc , glucocorticosteriods , interferons , opi in which hydrophobic active ( s ) may be dissolved . Generally , oids , infliximab , etanercept, adalimumab , cucumin , cat the dispersed phase may provide a hydrophobic environment echins , mycophenolate mofetil , an NO donor , a steroid ( e . g . either in a hydrophobic material or within the hydrophobic is not budesonide or another corticosteroid and is not an part of a micelle or promicelle . The dispersed phase may adrenal steroid , e . g . is not prednisone or a hydrocortisone , comprise a water- immiscible liquid . The water- immiscible administered alone or in combination with a xanthine or liquid may comprise a liquid lipid and optionally a solvent methylxanthine compound ) ; a cytokine e . g . is not sulfasal miscible therewith , in which solvent the water - insoluble izine (salicyl - azo -sulfapyridine , or “ SASP ” ) or a 5 -amin active ingredient is soluble . The ratio of liquid lipid to osalicylic acid product. The invention also comprises prod non - ionic surfactant may be in the range 1 - 4 : 1 by weight, ucts methods and uses which comprise at least one active optionally 1 . 2 - 3 . 0 : 1 by weight. The liquid lipid may be a mentioned in this paragraph . medium chain triglyceride (MCT ) composition , the medium [0180 ] The formulations may comprise gelatin as the chain triglyceride( s ) being one or more triglycerides of at water- soluble polymer . The gelatin may be substantially the least one fatty acid selected from C6 - C , , fatty acids. It will only water - soluble polymer . be understood that commercially available MCT composi [0181 ] The water - soluble matrix material may be selected tions useful in the invention are mixtures derived from from a hydrocolloid , a non -hydrocolloid gum and chitosan natural products and usually or always contain minor and derivatives thereof. amounts of compounds which are not MCTs; the term [0182 ] The formulations may comprise a unit solid which " medium chain triglyceride composition ” is therefore to be may be a minibead having a diameter of not more than 10 interpreted to include such compositions. mm , e . g . of notmore than 5 mm , the composition optionally [0188 ] The liquid lipid may be a caprylic / capric triglyc comprising a plurality of said minibeads . The minibead may eride , i .e . a caprylic / capric triglyceride composition (which be monolithic , optionally with layers thereon . The one or it will be understood may contain minor amounts of com more minibeads may comprise a controlled - release polymer , e . g . incorporated in the matrix and / or coated on it. The pounds which are not caprylic / capric triglycerides ). minibeads may comprise plural controlled release polymers , 10189 ]. For all embodiments of the invention , a water which may be present as a mixture or be separated , e . g . a insoluble active ingredient may have a solubility in the first controlled - release polymer may be comprised in a coat water- immiscible liquid of at least 5 mg/ ml , and often of at and a second (different ) controlled - release polymer may be least 10 mg/ ml , e . g . at least 25 mg/ ml , for example at least comprised in the matrix . The or each polymer may be 50 mg/ ml . associated with one or more excipients , e . g . a pore former . [0190 ] Said solvent which is optionally included in a The or each controlled -release polymer may be an extended water- immiscible liquid may be miscible with both the release polymer or an enteric polymer. The minibead ( s ) may liquid lipid and with water, e . g . it may be 2 - ( 2 - ethoxy ) have a coat which comprises the controlled release polymer ethanol. and optionally a polymer susceptible of degradation by [0191 ] The dispersed phase, e . g . water -immiscible phase bacterial enzymes. (water - immiscible droplets ) , may represent from 10 -85 % by [ 0183] In embodiments , the or each minibead comprises a dry weight of the composition . controlled - release polymer which is ethylcellulose com [0192 ] The unit solid or minibead may have a low water prised in a coating on the minibead and optionally in content. association with an emulsification agent, for example [0193 ] In an embodiment the pharmaceutical formulation ammonium oleate . The ethylcellulose may also be in asso is a capsule comprising a population of minibeads which ciation with a plasticizer, e . g . dibutyl sebacate or medium have a diameter of at most 10 mm and which comprise a chain triglycerides. The coating may further comprise poly surfactant- containing water -soluble polymer matrix material mer susceptible of degradation by bacterial enzymes . The and a coating on the matrix material, wherein the hydro polymer susceptible of degradation by bacterial enzymes philic surfactant has an HLB value of at least 15 , and may be water - soluble , preferably pectin . wherein the coating comprises a controlled -release polymer, [0184 ] The hydrophilic surfactant may have an HLB value optionally wherein the coating is a barrier membrane for of at least 15, and optionally of at least 18, e . g . of at least 20 extended release of the active agent ( s ) and / or is a coating or at least 25 . which resists becoming degraded or becoming of increased 10185 ). The hydrophilic surfactant may be an anionic sur permeability in the conditions of the GI tract above the colon factant. The anionic surfactant may have an HLB value of at but which becomes degraded or of increased permeability in least 30 , e . g . at least 35 , for example of 40 + 2 . The anionic the conditions of the colon . The minibeads may further surfactant may comprise or be an alkyl sulfate salt. The alkyl comprise in the polymer matrix part a non - ionic surfactant sulfate salt may be sodium dodecyl sulfate ( SDS ) . The comprising a poly (oxyethylene ) group and the hydrophilic water- soluble polymer matrix material may further contain surfactant may be an anionic surfactant. US 2018 / 0042989 A1 Feb . 15 , 2018 14
[0194 ] For all embodiments of the invention , the compo [0209 ] The formulating may comprise ejecting the emul sition may further comprise another active pharmaceutical sion through a single -orifice nozzle , e .g . having a diameter ingredient, in addition to said active agent( s ) . of from 0 . 5 - 5 mm , to form drops which are then caused or [ 0195 ] The composition may comprise a gelatin or other allowed to fall into a cooling oil or other hardening medium capsule containing a plurality of minibeads into which the and allowed to harden to form minibeads, after which the water -soluble polymer matrix material is formed . minibeads are recovered from the cooling oil and dried . [0196 ] Included in the invention is an intermediate product [0210 ] All optional features previously described in rela for use in manufacturing a composition of the disclosure, the tion to the invention are applicable to the below described intermediate product being a water - soluble polymer matrix methods and all other aspects and embodiments of the material formulated as a minibead having a diameter of no invention . Likewise , optional features described in relation more than 10 mm , e . g . of not more than 5 mm , in which to the below described methods are applicable to embodi matrix material an active agent is present in solution in the ments and aspects of the invention described earlier and late matrix itself or in a dispersed phase contained by the matrix . in this specification . [0197 ] Also disclosed is a method ofmaking dried emul sion formulations of the disclosure , which method com Surfactants prises mixing an oil phase with an aqueous phase compris [0211 ] In the description and claims of this specification , ing a water soluble polymer matrix material to form an the term “ surfactant” is employed as a contraction for emulsion and then causing the emulsion to solidify. The “ surface active agent” . For the purposes of this description emulsion may be formed into droplets which are then and claims, it is assumed that there are four major classifi exposed to a solidification medium ( e . g . a water immiscible cations of surfactants : anionic , cationic, non - ionic , and oil ) . In the case of a dried micelle composition , the oil phase amphoteric ( zwitterionic ) . The non -ionic surfactant remains is replaced by a surfactant phase . whole , has no charge in aqueous solutions, and does not [0198 ] The invention includes a colloidal composition , dissociate into positive and negative ions . Anionic surfac e .g . an emulsion or aqueous micelle composition , useful in tants are water - soluble , have a negative charge and disso making the formulations of the invention and comprising a ciate into positive and negative ions when placed in water . said active agent in solution in a phase of the colloid . The negative charge lowers the surface tension of water and 10199 ]. Further provided is an emulsion for use in manu acts as the surface - active agent . Cationic surfactants have a facturing an intermediate product of the disclosure , the positive charge , and also dissociate into positive and nega emulsion comprising oil droplets dispersed in an aqueous tive ions when placed in water . In this case , the positive ions phase characterised in that the aqueous phase comprises a lower the surface tension of the water and act as the water - soluble polymer matrix material and in that the emul surfactant. The amphoteric ( zwitterionic ) surfactant assumes sion comprises a said active agent in solution . In the case of a positive charge in acidic solutions and performs as a a micelle composition , the oil droplets are replaced by cationic surfactant, or it assumes a negative charge in an micelles . alkaline solution and acts as an anionic surfactant . [0200 ] The invention further includes a pharmaceutical [0212 ] Surfactants can also be classified according to their composition for oral administration , obtainable by : hydrophilic - lipophilic balance (HLB ) which is a measure of [0201 ] ( A ) mixing together at least the following mate the degree to which the surfactant is hydrophilic or lipo rials to form a colloid : philic , determined by calculating values for the different [ 0202 ] V ) a said active agent, which may be soluble regions of the molecule , as described ( originally for non or insoluble in water ; ionic surfactants ) by Griffin in 1949 and 1954 and later by [ 0203 ] vi ) an aqueous phase comprising water and a Davies . The methods apply a formula to the molecular water- soluble polymer material; weight of the whole molecule and of the hydrophilic and [0204 ] vii ) a hydrophobic liquid or a micelle- forming lipophilic portions to give an arbitrary ( semi- empirical) surfactant; scale up to 40 although the usual range is between 0 and 20 . [ 0205 ] viii ) optionally a hydrophilic surfactant hav An HLB value of 0 corresponds to a completely hydropho ing an HLB value of at least 10 ; bic molecule , and a value of 20 would correspond to a [0206 ] ix ) optionally one or more excipients which molecule made up completely of hydrophilic components . are miscible with or soluble in hydrophobic liquid or The HLB value can be used to predict the surfactant prop micelle - forming surfactant to increase the solubility erties of a molecule : of the immunosuppressant in said liquid or surfac tant, HLB Value Expected properties wherein any water- insoluble active ingredient is soluble in the hydrophobic liquid or micelle - forming surfactant when 0 to 3 antifoaming agent from 4 to 6 W / O emulsifier combined with any said one or more excipients ; and from 7 to 9 wetting agent [0207 ] (B ) formulating the colloid into a pharmaceuti from 8 to 18 an O / W emulsifier cal composition comprising a unit solid which com from 13 to 15 typical of detergents prises the colloid in a dry state . The composition may 10 to 18 solubiliser or hydrotrope be adapted to release the active ingredient at least into the colon . (0213 ] Although HLB numbers are assigned to surfactants [ 0208 ] The mixing together may comprise forming a clear other than the non - ionic , for which the system was invented , solution of the active ingredient in the hydrophobic liquid or HLB numbers for anionic , cationic , non - ionic , and ampho micelle - forming surfactant together with any said one or teric ( zwitterionic ) surfactants can have less significance and more excipients. often represent a relative or comparative number and not the US 2018 / 0042989 A1 Feb . 15 , 2018 15 result of a mathematical calculation . This is why it is of the other excipients and of the active principle (s ). Higher possible to have surfactants above the " maximum " of 20 . alkyl may in particular implementations of the invention be HLB numbers can however be useful to describe the HLB Cu - C . , or C . - C . , alkyl, and in some embodiments has a requirement of a desired application for a given emulsion length of no greater than C18 . system in order to achieve good performance . [ 0221 ] Instead of ( or as complement to ) the surfactant in the aqueous phase , the invention also contemplates use of Hydrophilic Surfactants for the Aqueous Phase surfactant - like emulsifiers ( also known as crystallisation [ 0214 ] In embodiments of the invention , the unit solid inhibitors ) such as, for example, HPMC (also known as comprises a hydrophilic surfactant which , without being hypromellose ) although their use is generally contemplated bound by theory , is believed at least partially to partition the in relatively smaller amounts to avoid high viscosity which aqueous phase (polymer matrix ) . may constrain processing options . [0215 ] Surfactants for such inclusion in the aqueous phase 0222 ] Other non - ionic surfactants which may be included of the inventive composition are preferably readily diffusing in the aqueous phase include poloxamers which are non or diffusible surfactants to facilitate manufacturing and ionic triblock copolymers composed of a central hydropho processing of the composition of the invention . The surfac bic chain of polyoxypropylene ( poly ( propylene oxide ) ) tantmay have an HLB of at least 10 and optionally of at least flanked by two hydrophilic chains of polyoxyethylene ( poly 15 , e . g . at least 30 and optionally of 38 -42 , e . g . 40 . Such (ethylene oxide ) ). Poloxamers are available commercially surfactants can be of any particular type ( cationic , anionic , under the trade name PluronicsTM . Such surfactants or non - ionic , zwitterionic ) and may comprise as a proportion of similar larger polymeric surfactants are aqueously soluble dry weight of the composition from 0 . 1 % to 6 % , e . g . 0 . 1 % and are therefore presented here as optional components of to 5 % . 0 . 1 % to 4 % or 0 . 1 % to 3 % , e . g . in a proportion of at the aqueous phase . However, they may be used to reduce the least 1 % and in particular between 1 . 0 and 4 . 5 or 5 % , for amount of or to replace a higher HLB polymeric component example within or just outside the 2 - 4 % range , for example of the oil phase ( see also separate section ) such as , for from 2 to 3 % or approximately 2 % or approximately 4 % . example , polyethoxylated castor oils (polyethylene glycol The invention includes formulations in which the hydro ethers) exemplified commercially as CremophorTM philic surfactant is , or comprises, an anionic surfactant, e . g . Diblock , tetrablock , multiblock , etc copolymers (poloxom a single anionic surfactant or a mixture thereof. ers ) are also included . [ 0216 ] Unless otherwise stated or required , all percentages [0223 ] Another type of polymeric aqueous soluble surfac and ratios are by weight. tant which may be used in a similar way are anionic 0217 ] Preferred anionic surfactants for inclusion in the copolymers based on methacrylic acid and methyl meth aqueous phase include perfluoro - octanoate (PFOA or PFO ) , acrylate in which the ratio of the free carboxyl groups to perfluoro - octanesulfonate (PFOS ) , sodium dodecyl sulfate ester groups is approx . 1 : 1 and with average molecular (SDS ), ammonium lauryl sulfate , and other alkyl sulfate weight is approx . 135 , 000 . Such a polymeric surfactant is salts , sodium laureth sulfate , also known as sodium lauryl available from Degussa under the trade name EUDRAGIT® ether sulfate (SLES ) and alkyl benzene sulfonate . A particu L 100 . lar class of surfactant comprises sulfate salts . A preferred [0224 ] The surfactant included in the aqueous phase is anionic surfactant in the aqueous phase is SDS . Mixtures of preferably present within ranges noted above . In the mini anionic surfactants are also contemplated . bead embodiment, avoidance of excess surfactant is desir [0218 ] The physical form of the surfactant at the point of able to avoid the “ golf ball effect” whereby minibeads when introduction into the aqueous phase during preparation plays dried have a plurality of point- sized dimples in their surface a role in the ease of manufacture of the composition accord (visible under the microscope ) . While not necessarily a ing to the invention . As such , although liquid surfactants can major concern , such dimples can lead to variability in be employed , it is preferred to utilize a surfactant which is coating if it is desired to apply for example a polymer coat in solid form ( e. g . crystalline , granules or powder ) at room to the minibeads. Although higher values within the pre temperature , particularly when the aqueous phase comprises ferred range generally increase the rate of egress /dissolution gelatin . of minibeads, the present inventors /applicants have found [0219 ] Possible non - ionic surfactants for the aqueous that in certain circumstances higher levels of surfactant phase include perfluorocarbons , polyoxyethyleneglycol included in the composition of the invention can cause a dodecyl ether ( e . g . Brij such as, for example , Brij 35 ) , Myrj counterintuitive drop in the in vitro dissolution profile ( e . g . Myrj 49 , 52 or 59 ) , Tween 20 or 80 ( also known as including a drop in the total amount dissolved of the Polysorbate ). Brij, Myrj and Tween products are available composition according to the invention . The concentration commercially from Croda . of surfactant above which the dissolution profile dropped ( or [ 0220 ] In general , mixtures of surfactants can be utilised total amount of dissolved composition dropped ) was e . g . to achieve optimum long term stability of the compo approximately 5 % by dry weight of the composition for sition of the invention with shorter chain surfactants in example when SDS is selected as the surfactant. In certain general facilitating shorter term stability (an aid to process embodiments , it is therefore preferred to have in the aqueous ing ) and longer chain surfactants facilitating longer term phase a surfactant , whether non - ionic or ionic , for example stability ( an aid to shelf life ) . In some embodiments , shorter anionic e . g . SDS , in an amount of less than 5 % by dry chain surfactants have up to Co alkyl ( e . g . Co Co- alkyl) as weight of the total composition ( for example , the composi the hydrophobic portion of the surfactant whilst longer chain tion may be in the form of beads or minibeads, wherein the surfactants have C , , or higher alkyl ( e . g . C1. - C . , alkyl) as aqueous phase contains SDS or another surfactant in an the hydrophobic portion of the surfactant. It is envisaged that amount of less than 5 % by dry weight of the beads/ C10 alkyl surfactants may facilitate processing or facilitate minibeads ) . In embodiments of the invention , the composi prolongation of shelf life , or both , depending on the identity tion , e. g. in the form of beads or minibeads, comprises in the US 2018 / 0042989 A1 Feb . 15 , 2018 16 aqueous phase surfactant in an amount of no more than 5 % , [0229 ] High surfactant concentrations in the dissolution no more than 4 . 5 % , no more than 4 % or no more than 3 % medium can generate very good in vitro data but which is by dry weight of the beads or minibeads . In one class of not necessarily predictive of in vivo performance ( e . g . embodiments , the surfactant is in an amount of at least 0 . 1 % pharmacokinetic profile ) . In contrast, incorporation of by dry weight of the beads or minibeads. In another class of (much lower overall quantities of) surfactant in one embodi embodiments , the surfactant is in an amount of at least 1 % ment of the minibeads of the invention produces unexpect by dry weight of the beads or minibeads. In a further class edly superior in - vivo performance . The inventors/ applicants of embodiments , the surfactant is in an amount of at least 2 % hypothesise (without wishing to be bound by the hypothesis ) by dry weight of the beads or minibeads . Higher levels of that surfactant in the dissolution medium is more playing the surfactant in the aqueous phase (e . g. above 5 % by weight of role of a dispersing agent (bringing other components into the total composition ) restrict the processing parameters for the dissolution medium ) rather than its classical role as an manufacturing when certain manufacturing approaches are aid to dissolution and that it is the surfactant included in the followed aqueous phase of this embodiment of the composition of the [0225 ] It is noteworthy that surfactants are used in disso invention which ensures or enables dissolution . In this lution testing media when complete dissolution of the com setting , the small amount of surfactant included in the position being studied is otherwise not achievable . In respect dissolution medium therefore makes the test more a disper of the amount of surfactant included in the aqueous phase of sion test than a dissolution test and achieves dissolution / the composition of the present invention as described above , dispersion maintenance for the purposes of compendial the inventors/ applications have surprisingly found that such methods . ( small ) quantities included in the composition have a much greater effect than larger quantities included in the dissolu Surfactants for the Water - Immiscible Phase tion medium . [0230 ] The oil phase , where present, may also include [ 0226 ] In the case of the minibead embodiment , the pres surfactant more hydrophobic than that chosen for the aque ent inventors hypothesise that the local concentration of ous phase , e . g . a non - ionic surfactant. The surfactant usually surfactant in and around the minibead as it dissolves or has an HLB value of at least 10 but, in any event, less than disperses is more effective than an otherwise greater con that of the hydrophilic surfactant. The non - ionic surfactant centration in the medium as a whole . It is also believed , typically comprises a poly ( oxyethylene ) group , e . g . com although the inventors /applicants do not necessarily intend prises a glycerol polyethylene glycol ricinoleate . to be bound by this or other hypotheses advanced in this text, [0231 ] Examples include polyethoxylated castor oils that the surfactant in the beads assists egress of active agent (polyethylene glycol ethers ) which can be prepared by from within the polymer coat ( if a coat is afterwards added reacting ethylene oxide with castor oil . Commercial prepa to the minibeads) and also possibly to shield the active agent rations may also be used as the surfactant e . g . those com from crystallisation and / or precipitation after release from mercial preparations which contain minor components such the bead . as, for example, polyethylene glycol esters of ricinoleic acid , [ 0227 ] In certain embodiments complete or substantially polyethylene glycols and polyethylene glycol ethers of glyc complete dissolution of active agent in USP / EP / JP etc erol. The preferred example is Cremophor by BASF Corp . dissolution apparatus using standard media can be achieved , also known as Cremophor EL . Alternative or additional using no or only minor amounts of surfactant in the disso surfactants include phospholipids such as, for example , lution medium , by incorporating in to the formulation of the phosphatidylcholine . In embodiments of the composition of invention ( e . g . dosage form ) one or more surfactants even the invention which comprise a phospholipid surfactant, the when the quantity of surfactant incorporated into the for phospholipid surfactant may be incorporated either in the mulation is much smaller than would have been required in aqueous phase or in the oil phase or both . If at least one the medium to achieve a comparable degree of dissolution of phospholipid surfactant is incorporated in each phase , it may a formulation containing no surfactant. The one or more be the same phospholipid surfactant in both phases or surfactants may be comprised in the aqueous phase ( the different in each . polymer matrix ) or the oil phase , or both , and are in [0232 ] The HLB of the surfactant for the water- immiscible particular comprised in at least the aqueous phase and phase, where present , may be from 10 - 20 , e . g . 10 - 15, and optionally also in the oil phase . optionally 11 - 20 (preferably 11 - 15 ) . [ 0228 ] These observations are particularly relevant to the class of minibead embodiments of the invention , in particu The Dispersed Phase : The Water - Immiscible Phase (Oil lar where an oil - soluble active agent is incorporated in an oil Phase ) phase and the minibead comprises a surfactant, e . g . in at [0233 ] Any pharmaceutically suitable oil may be used to least the aqueous phase ( polymer matrix ). On full dissolu constitute the oil phase (oil drops ) according to the inven tion of the composition of the invention in standard 900 tion . In terms of dry weight of the composition of the 1000 mL dissolution pots using compendial medium , the invention , the oil phase generally comprises a proportion concentration of surfactant in an exemplary embodiment from 10 % to 85 % , preferably 15 % to 50 % , more preferably would be of the order of 0 . 001 % i .e . much lower than the 20 % to 30 % or from 35 % to 45 % e . g . for vaccine formu amount ( around 0 . 5 % - 1 % ) typically added to the dissolution lations . The term " oil " means any substance that is wholly medium . Putting it another way , very significantly greater or partially liquid at ambient temperature or close - to - ambi amounts of surfactant would need to be included in this ent temperature e . g . between 10° C . and 40° C . or between embodiment of the composition of the invention in order to 15° C . and 35° C ., and which is hydrophobic but soluble in achieve a fully diluted equivalent concentration of surfactant at least one organic solvent. Oils include vegetable oils ( e . g . typically used in 900 - 1000 mL dissolution pots . neem oil ) , petrochemical oils , and volatile essential oils . The US 2018 / 0042989 A1 Feb . 15 , 2018 17 water- immiscible phase in particular comprises a liquid rogolglycerides (polyoxylglycerides ) , caprylocaproyl mac lipid , e . g . a liquid composition comprising triglycerides rogolglycerides and caprylic /capric triglyceride. In terms of and /or diglycerides, for example medium chain (C6 , C7, Cg, commercial products , particularly preferred oils in the lower Cg, C9, C10 , Cu or C12 ) diglycerides or triglycerides or HLB range are LabrafacTM Lipophile ( e . g . 1349 WL ) , combinations thereof. Labrafil, Labrasol, Captex 355 and Miglyol 810 . [0234 ] Oils which may be included in the oil phase include [0241 ] Particularly preferred surfactants in the higher poly - unsaturated fatty acids such as, for example, omega - 3 HLB category include polyethoxylated castor oils (polyeth oils for example eicosapentanoic acid (EPA ) , docosohexae ylene glycol ethers ) . The preferred commercial product for noic acid (DHA ), alpha - linoleic acid ( ALA ), conjugated example is Cremophor. linoleic acid ( CLA ) . Preferably ultrapure EPA , DHA or ALA 0242 ] While higher HLB surfactants can be considered or CLA are used e . g . purity up to or above 98 % . Omega oils surfactants , the invention also contemplates, additionally or may be sourced e . g . from any appropriate plant e . g . sacha inchi. Such oils may be used singly e . g . EPA or DHA or ALA alternatively , inclusion of any other appropriate ( non - ionic or CLA or in any combination . Combinations of such or other ) surfactant in the oil phase. components including binary , tertiary etc combinations in [0243 ] For certain active principles, particularly hydro any ratio are also contemplated e . g . a binary mixture of EPA phobic / lipophilic agents such as cyclosporin A for example , and DHA in a ratio of 1: 5 available commercially under the the present inventors / applicants have observed to their sur trade name Epax 6000 . prise that incorporation into the oil phase of a surfactant of [ 0235 ] Oils which may be included in the oil phase are high HLB and an oil of low HLB in a ratio of 1 - 4 : 1 by particularly natural triglyceride -based oils which include weight , e . g . 1 . 2 - 3 . 0 : 1 by weight, preferably 1 . 5 - 2 . 5 : 1 by olive oil, sesame oil, coconut oil, palm kernel oil. Oils which weight and most preferably 1. 8 -2 .2 : 1 by weight (high HLB : are particularly preferred include saturated coconut and low HLB ) advantageously stabilizes the emulsion before palm kernel oil- derived caprylic and capric fatty acids and and after immobilization of the oil droplets in the aqueous glycerin e .g . as supplied under the trade name MiglyolTM a phase . In this context “ stabilize ” means in particular that the range of which are available and from which one or more embodiment improves dissolution and /or dispersion of the components of the oil phase of the invention may be selected composition in vitro . including MiglyolTM 810, 812 ( caprylic / capric triglyceride ); [ 0244 ] By " high " HLB in this context is generally MiglyolTM 818 : (caprylic /capric / linoleic triglyceride ); Mig intended above 10 , preferably from 10 -16 , e . g . from 12 and lyolTM 829 : ( caprylic /capric / succinic triglyceride ; Mig 16 or 12 to 14 . By “ low ” HLB is generally intended below lyolTM 840 : (propylene glycol dicaprylate /dicaprate ). Note 10 , preferably in the range 1 to 4 , more preferably 1 to 2 . that MiglyolTM 810 /812 differ only in Cg/ C10 - ratio and [0245 ] The oil phase preferably also comprises a solvent, because of its low C10 - content, the viscosity and cloud point miscible with the oil, for the active principle . The oil phase of MiglyolTM 810 are lower . The MiglyolTM range is avail may therefore comprise a liquid lipid and a solvent miscible able commercially from Sasol Industries. As noted above , therewith , in which solvent the water - insoluble active ingre oils which may be included in the oil phase need not dient is soluble . The solvent for the active principle may be necessarily be liquid or fully liquid at room temperature . miscible with both the liquid lipid and with water. Waxy - type oils are also possible : these are liquid at manu 102461 Examples of suitable solvents are 2 -( 2 -ethoxy facturing temperatures but solid or semi- solid at normal ethoxyethanol available commercially under trade names ambient temperatures. Carbito1TM , Carbitol cellosolve , Transcuto1TM , Dioxito1TM , [0236 ] Alternative or additional oils which may be Poly - solv DETM , and Dowanal DETM ; or the purer Transcu included in the oil phase according to the invention are tolTM HP ( 99 . 9 ) . Transcutol P or HP, which are available medium chain triglyceride compositions such as for example commercially from Gattefosse , are preferred . Another pos LabrafacTM Lipophile manufactured by Gattefosse in par sible co - solvent is poly ( ethylene glycol) . PEGs ofmolecular ticular product number WL1349 . MiglyolTM 810 , 812 are weight 190 -210 ( e . g . PEG 200 ) or 380 -420 ( e . g . PEG 400 ) also medium chain triglyceride compositions. The medium are preferred in this embodiment. Suitable PEGs can be chain triglyceride( s ) mentioned herein are those which com obtained commercially under the name “ Carbowax ” manu prise one or more triglycerides of at least one fatty acid factured by Union Carbide Corporation although many selected from fatty acids having 6 , 7 , 8 , 9 , 10 , 11 or 12 alternative manufacturers or suppliers are possible . carbon atoms, e . g . C8 -C10 fatty acids . [0247 ] A particularly preferred oil phase according to the [0237 ] Other possible ( alternative or additional) oils invention is made up of an oil ( low HLB ) , a surfactant ( high include linoleoyl macrogolglycerides (polyoxylglycerides ) HLB ) and a solvent for the active principle . The oil may be such as , for example , Labrafil ( e. g . product number a liquid lipid e . g . an MCT composition . For example the M2125CS by Gattefosse ) and caprylocaproylmacrogolglyc following three commercial products : Transcutol P ( as sol erides such as, for example , Labrasol by Gattefosse . vent) , Miglyol 810 (as oil) and Cremophor e . g . Cremophor [ 0238 ] In one embodiment of the invention , the oil phase EL ( as surfactant) is particularly preferred . Miglyol 810 has comprises more than one component. For example , as just a low HLB and Cremophor has a high HLB . This particu mentioned , the oil phase may comprise a surfactant. larly preferred oil phase is preferably used to prepare ( and is [0239 ] Within this preferred embodiment, it is further preferably a component of ) a composition of the invention preferred that the HLB of the oil be in the range 0 - 10 comprising cyclosporin . In one embodiment , the composi (optionally 1 - 8 , e . g. 1 -6 and sometimes 1- 5 ) and the HLB of tion comprises an oil - soluble or hydrophobic antioxidant the surfactant be in the range 10 - 20 and optionally 11 - 20 e . g . hydralazine or BHT or carnosic acid or vitamin E . ( preferably 11 - 15 ) . [0248 ] The oil phase may also be a water - in - oil ( w / o ) [0240 ] Particularly preferred oils in the lower HLB cat emulsion so that the composition of the invention becomes egory include medium chain triglycerides , linoleoyl mac a water- in -oil - in - water ( w / o / w ) emulsion . US 2018 / 0042989 A1 Feb . 15 , 2018 18
[ 0249] The oil phase may include a said active agent known within the art . Although the size of the micelles is and /or one or more active principles and may also include given as a diameter this does not imply that the micelles one or more volatile or non - volatile solvents , which may be must be purely spherical species only that they may possess the same or different from the solvent or oil phase surfactant some approximately circular dimension . previously mentioned . Such solvents may for example [0255 ] The surfactant may be a non - ionic surfactant. The remain in the composition of the invention following pro surfactant may be a polyoxyethylated surfactant. The sur cessing e . g . initial dissolution of the active principle, and factant has a hydrophilic head which may be a hydrophilic have no particular function in the final composition . Alter chain , for example a polyoxyethylene chain or a polyhy natively, such solvents if present may function to maintain droxylated chain . the active principle in a dissolved state ( in solution ) within [0256 ] The surfactant of course has a hydrophobic part the oil phase or to facilitate dispersion , egress etc . In other and in particular a hydrophobic chain . The hydrophobic embodiments , the solvent may have partly or fully evapo chain may be a hydrocarbon chain , for example having at rated during processing and therefore be present in only least 6 carbon atoms and optionally at least 10 carbon atoms, minor quantities if at all . In a related embodiment , the and particularly of at least 12 carbon atoms; some hydro solvent, particularly when a solvent which is both oil and carbon chains have no more than 22 carbon atoms , for water -soluble is used , may be partly or completely present in example C10 -C20 , C12 - C20 or C15 - C20 hydrocarbon chains. It the aqueous phase of the composition according to the may be an alkyl chain , e . g . having a number of carbon atoms invention . An example of such a solvent is ethanol. Another just mentioned . It may be an alkenyl chain comprising one example is Transcutol which is already mentioned as a or more carbon - carbon double bonds, e . g . having a number co - solvent . of carbon atoms just mentioned . The surfactant may com [0250 ] It will be appreciated , therefore , that the invention prise a hydrocarbon chain , e . g . alkyl chain or alkenyl chain , provides inter alia a bead or minibead comprising a water that is substituted provided that it maintains a hydrophobic soluble polymer matrix material in which are dispersed characteristic . There may for example be one or two sub droplets of oil , the composition comprising an active prin stituents , for example a single substituent , e . g . selected from ciple and the oil comprising a combination of a high HLB halogen ( e . g . F or CI) , hydroxy , thiol oxo , nitro , cyano ; compound , e . g . a surfactant, and a low HLB compound , e . g . hydroxy or thiol substituents may be esterified by for an oil , and optionally including a co - solvent. example a fatty acid . One class of surfactants comprise a (0251 ] The oil droplets in the aqueous phase in its wet hydrocarbon monosubstituted by hydroxy ; optionally , at state during manufacture may be small enough (e . g . < 100 least a portion of the hydroxy groups of an aliquot of nm ) not to refract light, hence forming a transparent disper surfactant, e . g . of the surfactant in a bead , may be esterified sion . This is termed a microemulsion , as is well known in the by a fatty acid or mono - hydroxy fatty acid as disclosed art. herein or etherified by a fatty alcohol for example having at least 6 carbon atoms and optionally at least 10 carbon atoms, The Dispersed Phase : The Micelle Phase and particularly of at least 12 carbon atoms; some hydro [ 0252 ] As an alternative to an oil or wax phase as carbon chains have no more than 22 carbon atoms, for described above, the dispersed phase of the colloidal for example C10 -C20 , C12 -C20 or C15 - C20 fatty alcohols . mulations of the invention may comprise micelles , vesicles, [0257 ] The hydrophobic chain may be part of an esterified liposomes or nanoparticles , or at least the structures which fatty acid RP - COOH or of an etherified or esterified fatty result from drying aqueous colloids of such types . The ether R ? — COH where Rl is the hydrophobic chain , e. g . as invention in particular includes formulations in which the mentioned in the preceding paragraph . The ester - forming or, dispersed phase is micellar, i. e . formed of micelles and / or as the case may be, ether - forming group will typically promicelles . The term " promicelle ” refers to a part of a comprise a hydrophilic chain . formulation which will form a micelle upon contact with water, e . g . gastrointestinal contents . [0258 ] As mentioned , the surfactant may have a hydro [ 0253 ] A micelle - forming surfactant is present as micelles philic chain and may be a non - ionic surfactant, and may dispersed within the hydrogel- forming polymer in a " wet ” satisfy both requirements . The hydrophilic chain may be a (not yet dried ) composition made as an intermediate in the poly (ethyleneglycol ) , also known as poly (oxyethylene ) or manufacturing process described herein . It is believed also macrogol. The hydrophilic chain may be of the formula to be present as micelles in the dried composition but O CH , CH , ) . — OR where n is 5 or 6 to 50 and R is observability of micelles or micelle - like structures in the H or alkyl, e . g . ethyl or methyl. The invention includes dried composition is not a requirement of the invention . It is implementations in which n is from 6 to 40 , e . g . from 6 to mentioned at this point that the presence of a surfactant in 35 . In some embodiments , n is from 6 to 25 and optionally micelle form does not require that the entire surfactant is from 8 to 25 or from 8 to 15 . In other embodiments , n is content of a composition is in micelle form as it is consid from 8 to 50 or from 8 to 40 , e . g . is from 10 to 50 , 10 to 40 ered more probable that a portion of the surfactant will be or 10 to 35 . In a particular embodiment, n is 15 . For all outside the micelles . Thus in the " wet ” composition , hydrophilic chains of the formula ( O CH2CH ) n whether the hydrogel- forming polymer is in the gel state or OR , in one class of embodiments R is H . the sol ( liquid ) state may comprise the micelle - forming [0259 ] The hydrophilic chain may be a polyhydroxylated surfactant at a concentration above the critical micelle chain ( for example a C5- C20 e . g . C5 - C10 chain ) , e . g . having concentration . a hydroxy group on the carbon atoms of the chain , for [ 0254 ] The diameter of the dispersed micelles may be example a glucamide . between 0 .5 nm and 200 nm , 1 nm and 50 nm , or 5 nm and [ 0260 ] The micelle - forming surfactant may comprise a 25 nm . The size of the micelles may be determined by combination of a hydrophobic chain as described above and dynamic light scattering or diffusion NMR techniques a hydrophilic chain as described above . It may therefore be , US 2018 / 0042989 A1 Feb . 15 , 2018 19 or comprise , a macrogol ester of a fatty acid as described preceding section of this specification . Alternatively , a sur herein or a macrogol ether of a fatty alcohol as described factant may be combined with one ormore other compounds herein . at least potentially able to form micelles with the surfactant, [0261 ] Micelle - forming surfactants comprising a hydro optionally selected from cationic lipids and glycolipids, phobic chain and a hydrophilic chain can be selected from amongst others . As an additional option , a composition may the group consisting of: macrogol esters ; macrogol ethers ; comprise a plurality of surfactants as mentioned in the diblock copolymers ; triblock copolymers ; and amphiphilic preceding section of this specification and one or more other polymers . In certain embodiments of the invention any compounds at least potentially able to form micelles with the combinations of the group are included within the invention . surfactant, optionally selected from cationic lipids and gly [0262 ] Examples ofmacrogol esters which are suitable for colipids , amongst others . use in the present invention are macrogol esters of fatty acids [0269 ] The invention therefore includes compositions as having at least 6 carbon atoms and optionally at least 10 described herein which comprise : carbon atoms, and particularly of at least 12 carbon atoms; ( 0270 ] two or more micelle - forming surfactants , e . g . some fatty acids have no more than 22 carbon atoms, for two or more surfactants having a hydrophobic chain example C10 - C20, C12 - C20 or C15 - C20 fatty acids. The fatty and a hydrophilic chain acids may be saturated or unsaturated but are in particular [ 0271 ] a compound , e . g . a single compound or two or saturated . To bementioned are macrogol 25 cetostearyl ether more compounds, selected from cationic lipids and (Cremophor® A25 ) ; macrogol 6 cetostearyl ether (Cremo glycolipids phor® A6 ) ; macrogol glycerol ricinoleate 35 ( Cremophor® (0272 two or more micelle - forming surfactants and a EL ); macrogol- glycerol hydroxystearate 40 ( Cremophor® compound , e . g . a single compound or two or more RH 40 ) ; macrogol - 15 -hydroxystearate ( Solutol® HS 15 ) . compounds , selected from cationic lipids and glycolip Examples of macrogol ethers which are suitable for use in ids the present invention are macrogol ethers of fatty alcohols having at least 6 carbon atoms and optionally at least 10 The Aqueous Phase carbon atoms, and particularly of at least 12 carbon atoms; [ 0273 ] The principal component of the aqueous phase of some fatty alcohols have no more than 22 carbon atoms, for the colloided formulations according to the invention (pref example C1o - C20 , C12 -C20 or C15 - C20 fatty alcohols . The erably between 20 % and 70 % , more preferably between fatty alcohols may be saturate or unsaturated but are in one 30 % and 60 % , still more preferably between 35 % and 55 % , embodiment saturated . by dry weight thereof) is a water - soluble polymer matrix [ 0263 ] Examples of amphiphilic polymers which are suit material although other components may also be included as able for use in the present invention are : alkyl glucamides; described below . The inclusion of too little of the water fatty alcohol poly ( ethoxyl) ates also known as polyethoxy soluble polymer matrix material can for certain active prin lated alkyl ethers ; poly ( ethoxyl) ated fatty acid esters (Myrj ciples lead to non - incorporation or leaching of the active out or Solutol) ; fatty amide polyethoxylate ; fatty amine ethoxy of the composition , particularly when in the form of mini late; alkylphenol ethoxylate ; polyethoxylated sorbitan esters beads . For certain embodiments, for example micellar com (polysorbates ) ; polyethoxylated glycerides; or poly - glycerol positions, e . g . comprising Solutol® , or those comprising a esters . retardant ( see below ) , it is preferred that the aqueous phase [0264 ] Examples of copolymers, which are suitable for comprise from 55 % and 65 % of the dry weight of the use in the present invention are : pluronics ( poloxamers ); composition . polyvinylpyrollidone - polyvinylacetate (Plasdone S630 ) ; [0274 ] In the case of non - colloidal formulations, a mini aminoalkyl methacrylate copolymer ( Eudragit EPO ) ; meth bead may comprise only an aqueous phase ( i . e . matrix acrylic acid - methyl methacrylate copolymer (Eudragit phase ) and optional coating ( s ) . S100 , L100 ) ; polycaprolactone -PEG ; polycaprolactone [02751 . While mixtures of water- soluble polymer matrix methoxy — PEG ; poly ( aspartic acid ) -PEG ; poly (benzyl - L materials are contemplated by the invention , preferably the glutamate )- PEG ; poly ( D , L -lactide ) methoxy - PEG ; poly composition of the present invention comprises a matrix (benzyl - L -aspartate -PEG , or poly (L - lysine )- PEG material which is substantially a single material or type of [ 0265 ] In a preferred embodiment the micelle - forming material among those described herein and /or a matrix surfactant cis a macrogol ester , more preferably a macrogol which can be solidified without inclusion of specific addi ester that conforms to the European Pharmacopoeia mono tional polymeric components in the aqueous phase . How graph number 2052 macrogol - 15 -hydroxystearate , such as ever , mixtures may be preferred to achieve certain perfor Solutol® HS 15 marketed by BASF . mance characteristics . Thus it may be desired to incorporate [0266 ] Suitable surfactants comprise those which during certain constraining or retarding substances ( retardants ) into manufacture combine with the aqueous phase ( including the water - soluble polymer matrix . In certain embodiments , hydrogel - forming polymer ) in an amount above their CMC such incorporation permits a coat (or coating ) to be dis to form a clear liquid . Solutol® HS 15 is such a surfactant. pensed with . In other embodiments where a constraining or [0267 ] In certain embodiments the weight ratio of the retarding agent is included into the water -soluble polymer micelle - forming surfactant to the antigen is from 10 : 1 to matrix , a coat ( or coating ) may be present and desirable . For 100 : 1 , optionally from 50 : 1 to 100 : 1 . In some embodiments , example, incorporation of a retarding agent which is the ratio is from 80 : 1 to 90 : 1 . In particular embodiments , the insoluble in acid milieu ( such as the stomach ) is selected to ratio is from 50 : 1 to 60 : 1 . prevent or retard release in the stomach and a coating may [0268 ] In particular embodiments , the compositions of the not be needed i. e . the composition may be free of a coat/ invention comprise a combination of micelle - forming com coating . Alternatively , incorporation of a retarding agent pounds . Such a combination ofmicelle - forming compounds which is soluble in acid media may be selected to retard may consist of two or more surfactants as mentioned in the release in the intestine distal to the stomach . Again a coating US 2018 / 0042989 A1 Feb . 15 , 2018 20 may not be needed i . e . the composition may be free of a gelatins are also contemplated . Hydrocolloids which may be coat / coating . However, a composition according to the used according to the invention include those derived from invention which incorporates a retarding agent soluble in natural sources such as, for example , carrageenan (extracted acid media may optionally be coated e . g . with an acid from seaweed ), gelatin (extracted from bovine, porcine, fish resistant polymer to achieve particular advantage. Such a or vegetal sources ), agar ( from seaweed ) and pectin ( ex composition is protected from ( complete ) gastric release ( or tracted from citrus peel, apple and other fruits ) . A non gastric release is retarded ) owing to the effect of the acid animal based hydrocolloid may be preferred for certain resistant polymer coat. Distal to the stomach , following loss applications e . g . administration to vegetarians or to indi of the coat, the acid -soluble agent retards release because the viduals not wishing to ingest animal products for religious or milieu of the small and large intestine is no longer acid . health reasons. In relation to the use of carrageenan , refer Retarding or constraining agents insoluble in acid milieu ence is made to US patent application 2006 /0029660 A1 include polymers whose solubility is pH -dependent i. e . (Fonkwe et al) , the entirety of which is incorporated herein soluble at higher pH . Such polymers are described in detail by reference . in the section below entitled “ Coating ” and such polymers [0278 ] The immobilized aqueous phase of the composi may be used either as coats / coatings or as retarding agents tion according to one embodiment of the invention is incorporated into the water - soluble polymer matrix . An preferably a gel i . e . a substantially dilute crosslinked system , example of a suitable retarding agent mentioned in the which exhibits no flow when in the steady - state . The internal section below entitled “ Coating ” is HPMCP ( hydroxy - pro network structure of the solidified aqueous phase may result pyl- methyl -cellulose -phthalate also known as hypromellose from physical or chemical bonds, as well as crystallites or phthalate ) which is used to prevent release in the gastric other junctions that remain intact within an extending fluid environment since it is soluble above pH 5 . 5 — see that e . g . water. section for other examples of polymers soluble in non - acid [0279 ] In an alternative preferred embodiment, the poly ( basic ) media . HPMCP may also be used as a pore - former . mer matrix is a non - hydrocolloid gum . Examples are the Retarding or constraining agents soluble in acid milieu cross - linked salts of alginic acid . For example , aqueous include polymers whose solubility is pH - dependent i . e . solutions of sodium alginate gums extracted from the walls soluble at lower pH . Such polymers include cationic poly of brown algae have the well known property of gelling mers such as for example copolymers based on dimethyl when exposed to di- and trivalent cations . A typical divalent aminoethyl methacrylate , butyl methacrylate , and methyl cation is calcium , often in the form of aqueous calcium methacrylate . An example of such a cationic co - polymer chloride solution . It is preferred in this embodiment that the which may be used according to the invention is Eudragit E . cross - linking or gelling have arisen through reaction with PO commercially available from Evonik Industries4 . such a multivalent cation , particularly calcium . [ 0276 ] In one embodiment , the water- soluble polymer [0280 ] In an alternative preferred embodiment, the poly matrix material may be of one or more of those selected mer matrix is chitosan which can exist in the form of biogels from gelatin , agar , a polyethylene glycol, starch , casein , with or without additives as described e . g. in U .S . Pat. No . chitosan , soya bean protein , safflower protein , alginates , 4 ,659 ,700 (Johnson & Johnson ); by Kumar Majeti N . V . gellan gum , carrageenan , xanthan gum , phthalated gelatin , Ravi in Reactive and Functional Polymers , 46 , 1 , 2000 ; and succinated gelatin , cellulosephthalate -acetate , oleoresin , by Paul et al. in ST . P . Pharma Science, 10 , 5 , 2000 the polyvinylacetate , hydroxypropylmethyl cellulose , polymeri entirety of all 3 of which is incorporated herein by reference . sates of acrylic or methacrylic esters and polyvinylacetate Chitosan derivatives e. g. thiolated entities are also contem phthalate and any derivative of any of the foregoing. Mix plated . tures of one or more water- soluble polymers comprising the [0281 ] In the embodiment in which gelatin is the polymer matrix are also contemplated . In specific embodiments matrix of the invention , reference is hereby made to " bloom binary or tertiary etc combinations of any of the above strength ” , a measure of the strength of a gel or gelatin substances are foreseen . An unexpected advantage of com developed in 1925 by O . T . Bloom . The test determines the bining certain water -soluble polymers to form the matrix is weight ( in grams) needed by a probe ( normally with a that it allows for a reduction in the total amount of water diameter of 0 . 5 inch ) to deflect the surface of the gel 4 mm soluble polymer employed . This may have cost advantages without breaking it. The result is expressed in Bloom or may allow greater loading of other materials such as, for ( grades ) and usually ranges between 30 and 300 Bloom . To example , one or more active principles. Inclusion of (addi perform the Bloom test on gelatin , a 6 .67 % gelatin solution tion of ) a second water - soluble polymer to form the matrix is kept for 17 - 18 hours at 10° C . prior to being tested . may also give more strength to the composition of the [0282 ] According to the invention , where gelatin is the invention e . g . beads. polymer matrix , it is preferred to use gelatin with bloom [ 0277 ] In a preferred embodiment, the polymer matrix strength between 200 and 300 , preferably between 210 and material is a hydrocolloid i . e . a colloid system wherein the 280 . colloid particles are dispersed in water and depending on the [0283 ] According to the invention , where gelatin is the quantity of water available can take on different states , e . g . , water- soluble polymer matrix material, the gelatin may be gel or sol ( liquid ) . It is preferred to use reversible hydro sourced by a variety of means . For example , it can be colloids ( e . g . agar, gelatin etc ) as opposed to irreversible obtained by the partial hydrolysis of collagenous material , ( single - state ) hydrocolloids . Thermotropic hydrocolloids such as the skin , white connective tissues, or bones of ( also known as thermoreversable hydrocolloids ) can exist in animals . Type A gelatin is derived mainly from porcine skins a gel and sol state , and alternate between states with the by acid processing , and exhibits an isoelectric point between addition or elimination of heat. Gelatin is a thermo -revers pH 7 and pH 9 , while Type B gelatin is derived from alkaline ible, rehydratable colloid and is particularly preferred . Gela processing of bones and animal (bovine ) skins and exhibits tin derivatives such as, for example, succinated or phthalated an isoelectric point between pH 4 . 7 and pH 5 .2 . Type A US 2018 / 0042989 A1 Feb . 15 , 2018
gelatin is somewhat preferred . Gelatin for use in the inven - phase. An example is hydroxy propyl/ methyl cellulose tion may also be derived from the skin of cold water fish . (HMC or HPMC , hypromellose etc ) which may play other Blends of Type A and Type B gelatins can be used in the roles such as , for example , emulsifier ( see above ) . In addi invention to obtain a gelatin with the requisite viscosity and tion , the aqueous phase may include some or all of a solvent bloom strength characteristics for minibead manufacture . used during processing to dissolve , or facilitate dissolution [0284 ] Commercially gelatin can be obtained from the of, an active principle e . g . an active principle comprised in Sigma Chemical Company, St. Louis , Mo . USA or from the oil phase . An example is ethanol ( see discussion above Nitta (http :/ / www .nitta - gelatin . com ) . on use of solvents in oil phase ). [ 0285 ] Lower temperature gelatin ( or gelatin derivatives [0293 ] The invention includes compositions comprising a or mixtures of gelatins with melting point reducers) or other solid phase comprising a water - soluble polymer matrix polymer matrices able to be solidified at lower temperatures material and an oil phase dispersed in the solid phase . ( e. g . sodium alginate described above ) are preferred for example when the active principle to be incorporated in the Non -Colloidal Formulations composition of the invention is temperature - labile or whose activity may be affected by exposure to higher temperatures . [0294 ] Where the or each active ingredient is water [ 0286 ] According to the invention , where gelatin is the soluble , a single - phase formulation may be used consisting polymer , the starting gelatin material is preferably modified of a water- soluble polymer matrix containing dissolved before manufacture to produce “ soft gelatin ” by the addition active ingredient( s ) and optional coating (s ) . The preceding of a plasticizer or softener to the gelatin to adjust the section headed “ Aqueous Phase ” applies to the matrix phase hardness of the composition of the invention . The addition of such single - phase formulations . The polymer matrix may of plasticizer achieves enhanced softness and flexibility as be formulated as minibeads by ejecting it through a beading may be desirable to optimise dissolution and / or further nozzle as described above . processing such as, for example , coating. Useful plasticizers of the present invention include glycerin ( 1 , 2 , 3 - propanet Shape, Size and Geometry of Dried Colloidal Formulations riol) , D -sorbitol ( D -glucitol ), sorbitol BP (a non -crystalliz 0295 ] The dried colloidal compositions ( i . e . those obtain ing sorbitol solution ) or an aqueous solution of D - sorbitol able by drying a colloid ) and dried single phase composi and sorbitans ( e . g . Andidriborb 85 /70 ) . Other or similar low tions can be formed into a limitless number of shapes and molecular weight polyols are also contemplated . Polyethyl sizes . In the section below describing the process for making ene glycol may also be used although this is less preferred the composition , various methods are given including pour and indeed particularly preferred compositions of the inven ing or introducing a fluid emulsion into a mould where it tion are free or substantially free of PEG or derivatives hardens or can be caused to harden . Thus the composition thereof. Glycerin and D - sorbitol may be obtained from the can be created in whichever form is desired by creating an Sigma Chemical Company, St. Louis ,Mo . USA or Roquette , appropriate mould ( e . g . in the shape of a disc , pill or tablet ) . France . However , it is not essential to use a mould . For example , the [ 0287] As noted above, some constituents of the present composition may be in the form of a sheet e . g . resulting from invention may play more than one role . For example when pouring a fluid emulsion onto a flat surface where it hardens one of the active principles ( see below ) is ibuprofen , it may or can be caused to harden . also act as a plasticiser owing to its particular physico [0296 ] Alternatively , the composition may be in the form chemical properties . Choice of ibuprofen has particular of spheres or spherical- like shapesmade as described below . advantages in relation to higher loading as " conventional ” Preferably , the composition of the invention is in the form of plasticiser, for example dibutyl sebacate or DBS , may be substantially spherical, seamless beads , especially mini reduced in quantity . Alternatively it is contemplated that the beads . The absence of seams on the minibead surface is an surfactants discussed above may be selected for their plas advantage e .g . in further processing , for example coating , ticiser characteristics to achieve particular advantage. since it allowsmore consistent coating , flowability etc . The [ 0288 ] Softeners , if utilized , can be ideally incorporated in absence of seams on the minibeads also enhances consis a proportion rising to 30 % , preferably up to 20 % and more tency of dissolution of the minibeads . preferably up to 10 % by dry weight of the composition of [0297 ] The preferred size or diameter range of minibeads the invention , even more preferably between 3 and 8 % , and according to the invention can be chosen to avoid retention most preferably between 4 % and 6 % . in the stomach upon oral administration of the minibeads . [ 0289 ] As noted in more detail above in the section on Larger dosage forms are retained for variable periods in the surfactants , it is preferred to include one or more surfactants stomach and pass the pyloric sphincter only with food in the aqueous phase . Certain surfactants may also act as whereas smaller particles pass the pylorus independently of plasticisers or softeners or vice versa . food . Selection of the appropriate size range ( see below ) [0290 ] Although not essential, the aqueous phase may also thus makes the prediction of therapeutic effect post - dosing optionally contain a disintegrant where it is particularly more accurate . Compared to a single large monolithic oral desired to enhance the rate of disintegration of the compo format such as, for example , a traditional compressed tablet, sition of the invention . a plurality of minibeads released into the GI tract (as [ 0291] Examples of disintegrants which may be included foreseen by the present invention ) permits greater intestinal are alginic acid , croscarmellose sodium , crospovidone, low lumen dispersion so enhancing absorption via exposure to substituted hydroxypropyl cellulose and sodium starch gly greater epithelial area , prevents irritation ( e . g . as otherwise colate . seen with NSAIDs) and achieves greater topical coating [0292 ] A crystallisation inhibitor (e . g . approximately 1 % ( e . g . as may be desired for local drug effect in certain parts by dry weight of the composition ) may also be included in of the GI tract for example the colon ) . Reduction of resi the composition of the invention , preferably in the aqueous dence time in the ileo -caecal junction is another advantage . US 2018 / 0042989 A1 Feb . 15 , 2018 22
[0298 ] The dried colloidal or non -colloidal composition is minibeads of this embodiment of the invention may readily preferably monolithic meaning internally ( i. e . cross - section be compressed into a capsule to adopt the inner form of ally ) homogeneous . This is particularly preferred for the whichever capsule or shell may be desired leaving much minibead embodiment. reduced , e . g . essentially no , dead / void space . Alternatively [ 0299] The minibeads mentioned herein generally range in the dead or void space can be used to advantage by sus diameter from 0 . 5 mm to 10 mm with the upper limit pending minibeads in a vehicle such as , for example , an oil preferably 5 mm , e . g. 2 .5 mm . A particularly convenient which may be inert or may have functional properties such upper limit is 2 mm with 1 . 7 mm being particularly pre as , for example , permeability enhancement or enhanced ferred . The lower limit can preferably be 1 mm , e . g . 1 . 2 mm , dissolution or may comprise an active ingredient being the morem preferably from 1 . 3 mm , most preferably from 1. 4 same or different from any active ingredients in the bead . mm . In one embodiment the diameter is from 0 . 5 to 2 . 5 mm , For example , hard gelatin capsules may be filled with a for example from 1 mm to 2 mm . liquid medium combined with uncoated and /or coated [ 0300 ] In embodiments, the minibeads are monodisperse . beads . The liquid medium may be or comprise one or more In other embodiments , the minibeads are not monodisperse . of the oil phase constituents described herein or it may be By “ monodisperse ” is meant that for a plurality of minibeads one or more surfactants . Particularly preferred but non ( e . g . at least 100 , more preferably at least 1000 ) the mini limiting examples are corn oil and the commercial products beads have a coefficient of variation (CV ) of their diameters known as Span 85 , Labrafac , Transcutol P and Tween 80 . of 35 % or less, optionally 25 % or less, for example 15 % or [ 0304 ] Another possible form of the dried colloidal com less , such as e . g . of 10 % or less and optionally of 8 % or less , positions is as a capsule in which the core of the composition e . g . 5 % or less . A particular class of polymer minibeads has is a solid ( e . g . gastro -retentive float material such as , for a CV of 25 % or less . CV when referred to in this specifi example , bicarbonate salts ) or a fluid ( a gas or a liquid ) . If cation is defined as 100 times ( standard deviation ) divided the core is a liquid , it may contain an active principle and / or by average where " average ” is mean particle diameter and excipients which may be the same or different from those standard deviation is standard deviation in particle size . described above . Like the hemispherical beads described Such a determination of CV is performable using a sieve . above, such capsules may have two halves of different [0301 ] The minibeads may have a CV of 35 % and a mean constitution and sealed hermetically to retain the internal diameter of 1 mm to 2 mm , e . g . 1 . 5 mm . The minibeads may fluid . An internal layer e . g . internal film layer of non have a CV of 20 % and a mean diameter of 1 mm to 2 mm , aqueous material on the inner face of the sphere, may be e. g. 1. 5 mm , e .g . a CV of 10 % and a mean diameter of 1 mm included if it is desired that the core be an aqueous liquid to 2 mm , e . g . 1 . 5 mm . In one class of embodiments , 90 % of such that the internal layer prevents the aqueous core from beads have a diameter of from 0 . 5 mm to 2 . 5 mm , e . g . of coming into contact with the inner surface of the capsule . from 1 mm to 2 mm . With or without an intermediate layer, the core may be a [ 0302 ] Another possible form of the composition is as variant of the dried colloidal compositions so that the hemispherical beads two of which may optionally be joined composition of the invention , in the minibead embodiment, at the flat face to create a single minibead with two distinct comprises a core made from a first composition according to halves , each having a distinct composition , if that is desired , the invention and a capsule made from a second composition e . g . each containing different active principles or the same according to the invention . active principles but different excipients e . g . to achieve [ 0305 ] The minibead embodiment of the invention , while differing permeability , solubilization or release profiles as by itself offering a range of solutions to the issues identified between the two hemispheres. above , may also be used as a starting point for creation of [0303 ] The embodiment in which the composition takes further e . g . pharmaceutical or forms for example by using the form of minibeads can be further developed to create a the minibead as a nonpareil seed on which additional layers larger mass of minibeads e. g . via compression (with appro of material can be applied as is well known to a person priate oil or powder - based binder and / or filler known to skilled in the art e . g . of pharmaceutical science . Thematerial persons skilled in the art of pharmaceutical formulation ) and of the additional layers may comprise the same or different with the option of including additional quantities of the same active principle and / or the same or different excipients as are active ingredient as in the composition of the invention or a described in this document. Such variants allow differential different active ingredient. For example , the composition of release of the same or different active principles and facili the invention may take the form of beads which comprise an tate inclusion of multiple fixed -dose combination products active agent or combination of active agents as disclosed as for example discussed in connection with the popularly herein and the binder or filler comprises MMF, mycophe termed “ polypill” which denotes a single pill comprising nolate mofetil , an immunosuppressant ) . A compressed mass more than one active principle in a fixed dose combination . of minibeads may disintegrate at a different rate in different [ 0306 ] The formulations, whether or not dried colloidal conditions than a unitary moulded form of the same shape . formulations, may have a coat of additional material on its The larger ( e . g . compressed ) mass may itself take a variety outer surface . This coat may be applied in a number of ways, of shapes including pill shapes, tablet shapes , capsule shapes including drug layering , as described more particularly in etc . A particular problem which this version of the minibead the section below entitled " coating ” . In one such embodi embodiment solves is the “ dead space ” (above the settled ment, the formulation comprises an acid within the formu particulate contents ) and / or “ void space ” (between the par lation , for example within a bead , e . g . included within the ticulate content elements ) typically found in hard gel cap water soluble polymer matrix or as a liquid core in minibead sules filled with powders or pellets . In such pellet - or format and bicarbonate applied as a coat e . g . by drug powder- filled capsules with dead / void space , a patient is layering . If the formulation , e . g . minibead , has a polymeric required to swallow a larger capsule than would be neces coat, e . g . to control release into the colon , the bicarbonate sary if the capsules contained no such dead space . The may optionally or additionally be included in or be absent US 2018 / 0042989 A1 Feb . 15 , 2018 22 from the coating polymer. This composition is intended to from a powder which generally derives from micron -sized release carbon dioxide in the GI tract e .g . to reduce pain or particles incorporating a single or a small number of oil to reduce inflammation . The formulation may comprise an droplets often following coalescence of smaller droplets acid to enhance the solubility of active principles of various during spray - drying . While powder embodiments are not pKa ( acid dissociation constant ) in the small intestine or excluded , the composition of the invention , if particulate , colon . Alternatively , the formulation may comprise a base to preferably comprises particles larger than powder particles enhance the solubility of active principles of various pKa in such that the composition is in a non -powdered form . the stomach . [0314 ] Where the formulation is in the form of minibeads , a plurality of minibeads may be presented in a single format Other Characteristics of Dried Colloidal and Non - Colloidal e .g . contained in a single hard gel capsule which releases the Formulations minibeads e . g . in the stomach . Alternatively the minibeads [0307 ] The colloidal compositions, in certain embodi may be presented in a sachet or other container which ments , comprises one or more elements , components , permits the minibeads to be sprinkled onto food or into a excipients , structural features , functional features or other drink or to be administered via a feeding tube for example aspects of the prior art described above . a naso - gastric tube or a duodenal feeding tube . Alternatively , [0308 ] To summarise a limited number of embodiments of the minibeads may be administered as a tablet for example the invention , the composition as described above and if a plurality of minibeads are compressed into a single tablet elsewhere herein may additionally be one or more of the as described elsewhere herein . Alternatively , the minibeads following : substantially water - free , in a gel state , in a solid may be filled e . g . compressed into a specialist bottle cap or state , undissolved , non -powdered , formed , shaped , and not otherwise fill a space in a specialised bottle cap or other in solution . element of a sealed container (or container to be sealed ) such [ 0309 ] Unless geometrically designed to comprise inner that e . g . on twisting the bottle cap , the minibeads are aqueous compartments ( e . g . w / o / w format or capsular for released into a fluid or other contents of the bottle or vial mat with liquid core ), it is desirable that the colloidal such that the beads are dispersed ( or dissolve ) with or formulations of the invention are essentially or substantially without agitation in such contents . An example is the Smart dry , e . g . contains less than 5 % , preferably less than 1 % of Delivery Cap manufactured by Humana Pharma Interna free water by weight. Minibeads are preferably homoge tional (HPI ) S . p . A ., Milan , Italy . A related or similar neous although processing conditions may be varied ( see approach is also contemplated for e . g . timed release of below ) to achieve for example heterogeneity such as, for minicapsules into a reactor, feeding environment e . g . tank , example , a harder skin and softer core with less than incubator etc . complete immobilization of oil droplets towards the core as [0315 ] The minibeads so - presented may be of a single opposed to the surface of the bead . Larger ( e . g . non -beaded ) type (or population ) or may be of multiple types ( or popu forms or shapes of the composition according to the inven lations ) differing between populations in relation to one or tion may particularly be engineered to embody such hetero more features described herein e . g . different API or different geneity . excipients or different physical geometry , coated, multiply [ 0310 ] The low free -water content is a distinguishing coated , uncoated etc . feature of certain embodiments of the colloidal compositions 10316 ] In one embodiment, the invention allows for mini i . e . dried colloidal compositions . The free -water content can beads having immediate release ( IR ) characteristics e . g . be measured using thermogravimetic analysis ( TGA ) , for bearing no coat, enteric - only coat or coat designed to example with commercially available instrumentation , e . g . prevent release and /or dissolution of the bead only for a using a TGA Q 500 of TA Q series instrument. TGA limited time or lacking a retardant in the aqueous phase . In measures changes in weight in relation to a change in another embodiment, the invention allows for minibeads temperature . For example , a TGA method can comprise a having delayed or sustained release (SR ) characteristics e . g . temperature scan , e. g . from 20 to 400° C . at 20° C . per bearing a coat (or more than one coat) as described in more minute , where the moisture content is obtained from the detail elsewhere herein , particularly in the section entitled sample weight loss at about 100 degrees Celsius . " coating " . The invention also provides for an embodiment in [ 0311 ] In one embodiment , the dispersed phase , e . g . oil which immediate release minibeads are produced in com droplets is homogeneously dispersed in the solidified aque bination with a Sustained Release or Controlled Release ous phase (or in some embodiments the water - soluble poly (CR ) minibeads in varying ratios of IR : SR /CR . The imme mer matrix material ) with substantial absence of coalescence diate release minibeads can be combined with a Sustained or between adjacent oil droplets . Thus the colloid is preferably Controlled release minibead component in the following maintained during solidification . Coalescence of neighbour ratios ( w / w by potency ) e . g . 10 % Immediate Release ( IR ) + ing oil droplets ormicelles , preferably only does so , if at all , 90 % Sustained (SR )/ Controlled Release (CR ) minibeads; on rehydration of the composition of the invention . 20 % IR + 80 % SR /CR ; 30 % IR + 70 % SR /CR ; 40 % IR + 60 % [ 0312 ] Depending on process parameters , oil droplet size SR /CR and 50 % IR + 50 % SR / CR . can vary broadly e . g . from 10 nm to 10 um ( diameter ) . However , the inventors / applicants have found that it is Other Formulation Formats beneficial to maintain droplet size in the range from 100 nm [0317 ] The invention is not limited to colloidal formula to 1 um , e .g . from 300 - 700 nm . The term " emulsion " tions and optionally coated matrix phase - only formulations therefore includes microemulsions and nanoemulsions . as extensively discussed herein . Insofar as any embodiment [ 0313 ] The colloidal compositions generally comprise permits , an alternative solid oral formulation may be used . multiple oil drops or droplets within a moulded or shaped For example , two -layer and three - layer seamless minibeads form e . g . a minibead which might typically contain many are contemplated , which may be formed by ejecting through hundreds or thousands of droplets or micelles as distinct a central orifice of a bead - forming nozzle a liquid core US 2018 / 0042989 A1 Feb . 15 , 2018 24 material and simultaneously ejecting surrounding layers FG4095 , AGN - 2979 , metirosine , 3 - iodotyrosine , through one or two orifices arranged concentrically around aquayamycin , bulbocapnine , oudenone, TM 6008 , TM the central orifice . The outer layer is typically a water 6089 , siRNAs against hydroxylases and antisense therapeu soluble polymer matrix as described herein . The internal tics against hydroxylases, e. g. against PHD1, and combina phase of a two- layer minibead may have the same consti tions thereof. In any event, two or more hydroxylase inhibi tution as a hydrophobic phase described elsewhere herein . tors may be used . [0318 ] Those compositions which comprise a macrolide F0325 ] In many cases , the products , methods and uses of immunosuppressant preferably include a hydrophobic phase the disclosure comprise or involve , or may comprise or in which the immunosuppressant is dissolved . The hydro involve , an immunosuppressant. The identity of the immu phobic phase may be a solution , therefore . nosuppressant is not critical. It may be , or comprise , any one [ 0319 ) Active agents which are more readily water or more of: a calcineurin inhibitor, cyclosporin A soluble, e . g . the hydroxylase inhibitor hydralazine , may be ( ciclosporin ); mTOR inhibitors , eg . sirolimus (rapamycin ), incorporated in a hydrophilic phase ( e . g . a water soluble sirolimus derivatives for example everolimus, 32 - deoxora polymer ) in dissolved or particulate form . pamycin ; a mycophenolate, eg .mycophenolic acid ; metho [ 0320 ] Solid dosage forms for oral administration include trexate ; azathioprine or mercaptopurine ; mitoxantrone ; capsules, minicapsules , beads, powders and granules. In cyclophosphamide ; macrolide immunosuppressant, angio such solid dosage forms, the active compound is typically tensin II inhibitors ( e . g . Valsartan , Telmisartan , Losartan , mixed with at least one inert , pharmaceutically acceptable Irbesatan , Azilsartan , Olmesartan , Candesartan , Eprosartan ) excipient or carrier such as sodium citrate or dicalcium and ACE inhibitors e . g . sulfhydryl- containing agents ( e . g . phosphate and / or one or more : a ) fillers or externders such Captopril, Zofenopril ) , dicarboxylate - containing agents as starches, lactose , sucrose , glucose , mannitol and silicic ( e . g . Enalapril , Ramipril, Quinapril, Perindopril , Lisinopril , acid ; b ) binders such as carboxymethylcellulose , alginates , Benazepril, Imidapril, Zofenopril , Trandolapril) , phosphate gelatin , polyvinylpyrrolidone, sucrose and acacia ; c ) humec containing agents ( e. g. Fosinopril) , casokinins, lactokinins tants such as glycerol; d ) disintegrating agents such as and lactotripeptides. agar - agar , calcium carbonate , potato or tapioca starch , alg [0326 ] Some aspects and implementations of the invention inic acid , certain silicates and sodium carbonate ; e ) solution involve an active agent selected from calcineurin inhibitors , retarding agents such as paraffin ; f) absorption accelerators macrolide antibiotics and mTOR inhibitors. such as quaternary ammonium compounds ; g ) wetting [0327 ] Exemplary calcineurin inhibitors are cyclosporins , agents such as cetyl alcohol and glycerol monostearate ; h ) tacrolimus , and pimecrolimus. absorbents such as kaolin and bentonite clay and i ) lubri [ 0328 ] For examples of mTOR inhibitors useful in the cants such as talc , calcium stearate , magnesium stearate , invention the reader is referred to WO2007 /068462 , which solid polyethylene glycols , sodium lauryl sulfate and mix is incorporated herein by reference in its entirety . Particular tures thereof. In the case of capsules , tablets and pills, the examples are rapamycin , 40 - 0 - ( 2 - hydroxy ) - ethyl- rapamy dosage form may also comprise buffering agents . Solid cin , 32 - deoxorapamycin , 40 - [3 -hydroxy -2 - hydroxy compositions of a similar type may also be employed as methyl ) - 2 -methylpropanoate ) - rapamycin , ABT578 and fillers in soft and hard - filled gelatin capsules using such AP23573 . excipients as lactose or milk sugar as well as high molecular [0329 ] As examples of macrolide immunosuppressants weight polyethylene glycol, for example . may be mentioned tacrolimus, ascomycins, sirolimus, [ 0321 ] Suitably, the formulations contain a dissolution aid . cyclosporin , pimecrolimus. The dissolution aid is not limited as to its identity so long as [0330 ] An active agent may be water soluble and dis it is pharmaceutically acceptable . Examples include non solved in a water - soluble polymer, e . g . a hydrogel- forming ionic surfactants ; ionic surfactants ; and amphoteric surfac polymer, comprised in the formulation . The polymer may tants . constitute the polymer matrix of a colloidal formulation as [0322 ] The solid dosage forms can be prepared with extensively described herein . coatings and shells as well known in the pharmaceutical [0331 ] An active agent may be hydrophobic and dissolved formulating art and described elsewhere herein . They may in a hydrophobic medium , e . g . an oil , a wax or the interior optionally contain opacifying agents and may also be of a of a micelle . The oil, wax or micelle may form the dispersed composition such that they release the active ingredient ( s ) phase of a colloidal formulation as extensively described only , or preferentially , in a certain part of the intestinal tract , herein . and/ or in delayed fashion . [0332 ] The active agent( s ) are usually in solution in a ( 0323 ] A solid dosage form may comprise a waxy phase in liquid ( e . g . an oil or a micelle ) or in a solid or semi- solid which one or more active agents are dissolved . Suitable medium ( e. g . a wax or a water- soluble polymer) . The second waxy materials are described elsewhere herein . aspect of the invention demands that its minibeads comprise an active agent in solution . It will be recalled in this respect Active Ingredients that an active agent may be dissolved in a liquid medium to [ 0324 ] The active agents which may be used in the various form a solution which is clear to the human eye prior to aspects of the invention are described under the heading manufacturing of the formulation , and the invention “ Brief Summary of the Disclosure ” and in the claims. In includes a formulation obtainable by, i . e . having the char many cases , the products ,methods and uses of the disclosure acteristics of a formulation obtained by, incorporating such comprise or involve , or may comprise or involve, a hydroxy a solution in a minibead or other solid formulation and lase inhibitor. The hydroxylase inhibitor may inhibit an optionally drying and cooling the minibead or other formu asparaginyl hydroxylase ; it may inhibit a prolyl hydroxy lation . lase; it may inhibit both . The hydroxylase inhibitor may be [0333 ] The minibeads of the disclosure provide a hydro selected from , or comprise , DMOG , hydralazine , FG -4497 , philic environment ( the water- soluble polymer) for hydro US 2018 / 0042989 A1 Feb . 15 , 2018 25 philic drugs and a hydrophobic environment (the dispersed limiters, essential oils such as, for example , omega 3 oils , phase ) for hydrophobic drugs . Hydrophobic drugs are typi natural plant extracts such as , for example , neem , ion cally in solution in the dispersed phase but suspensions are exchange resins , bacteria degradable conjugation linkers not excluded . Hydrophilic drugs are often also in solution , in such as, for example , azo bonds, polysaccharides such as, the matrix phase , but may be particulate, e . g . as nanopar for example, amylose , guar gum , pectin , chitosan , inulin , ticles, in the matrix phase . cyclodextrins, chondroitin sulphate , dextrans, guar gum and [0334 ] The active agents may be used in combination locust bean gum , nuclear factor kappa B inhibitors , acids therapy with any other active pharmaceutical ingredient. For such as, for example, fumeric acid , citric acid and others, as example the other ingredient may be an anti - fibrotic agent, well as modifications thereof. for example selected from caspase inhibitors, peroxisome [0341 ] The composition may further comprise excipients proliferator - activated receptor - g (PPAR - g ) agonists such as to reduce systemic side effects associated with absorption in pioglitazone , TGF- b blockers , colchicines, relaxin , adi the small intestine including , but not limited to , antioxidants , ponectin , endothelin A , angiotensin receptor blockers , can such as , for example , curcuminoids , flavanoids or more nabinoids and agents altering the MMP- TIMP balance , and specifically including curcumin , beta - carotene , a - tocoph wound healing agent ( Ilodecakin , Mannose - 6 - Phosphate ) . erol, ascorbate or lazaroid . This is particularly the case in the context of fibrotic 0342 ] The composition may further or separately com disorders of the GIT and their treatment as described herein . prise antioxidants ( such as , for example , ascorbic acid or [ 0335 ] Where two or more actives are use in combination BHT - butyl hydroxy toluene ) taste -masking or photosensi therapy they may be in a combination formulation , e . g . two tive components or photoprotective components . Antioxi or more actives may be included in a single population of dants may be incorporated in the aqueous phase ( e . g . hydro minibeads or a composition ( e . g . capsule or other container ) philic antioxidants ) or in the oil phase ( e . g . hydrophobic may comprise two or more different populations of mini antioxidants such as , for example , vitamin E ) for example up bead , the minibeads of each population having one or more to 1 % by weight, preferably between 0 .01 and 0 .50 % by actives not found in the other population . Alternatively , any weight, more preferably between 0 .10 to 0 .20 % by weight. two actives which are co - administered may be administered in separate formulations , e . g . simultaneously , separately or Process for Making Colloidal Formulations sequentially and often simultaneously . [ 0343 ] The reader is notified that it is important to refer to [0336 ] As noted above, more than one active principle this section in relation to the Examples . may be incorporated in a single minibead and / or in distinct [0344 ] A basic method for making colloidal formulations populations of minibeads within a single dosage form , e . g . is to mix a fluid form (preferably a solution ) of the polymer hard gel capsule . The composition of the invention lends (or mixture of polymers ) chosen to be the water -soluble itself to fixed dose combinations of particular drugs , e . g . polymer matrix material ( e . g . gelatin , gum , alginate etc as comprising an active agent as described herein with acetyl described more generally elsewhere herein and in any event salicylic acid (ASA ) optionally in admixture with other components described [ 0337 ) The present invention also provides methods of above ) with a dispersed phase material, e . g . a surfactant treatment of one or more of the above diseases using the phase or an oil phase , to form homogeneous fluid emulsion . composition described herein . Taking account of the final composition required ( as described elsewhere herein ), the dispersed phase and the Other Active Excipients aqueous phase may be mixed in a proportion in the range [0338 ] The heading of this section is for convenience only 1 : 6 - 10 , particularly approximately 1 : 7 or 1 : 8 for an oily and does not imply strict categorisation . For example , a disperse phase or 1: 1 to 1: 4 for a surfactant (micellar ) category , substance or active principle described within this dispersed phase . In general, only gentle stirring of the " other active excipients ” may also be considered to fall components is required using a magnetic or mechanical within another section or category in this patent application . system e . g . overhead stirrer as would be familiar to a person One (non - limiting ) example is the group of substances skilled in the art to achieve emulsification . Continuous known as phospholipids which , according to the invention stirring is preferred . Any appropriate laboratory stirring may be excipients, permeability enhancers or active prin apparatus or industrial scale mixer may be utilized for this ciples ( e . g . phosphatidylcholine which is useful for instance purpose for example the Magnetic Stirrer (manufactured by in the treatment of inflammatory bowel disease ) . Stuart) or Overhead Stirrer (by KNF or Fisher ) . It is pre 10339 ] However, in general terms, the invention foresees ferred to set up the equipment in such a way as to minimise optional incorporation into the formulation of one or more of evaporation of contents such as, for example , water. In one the following substances or categories of substances in embodiment of the process of the invention , it is preferred addition to the primary active agent. For example , the to utilise a closed system for stirring in order to achieve this composition may contain a protectant such as , for example , aim . a proteolytic enzyme inhibitor or a protector against acid [0345 ] Where the polymer matrix is substantially consti degradation or both ( e . g . an alkali for example sodium tuted by gelatin with the addition of sorbitol, the aqueous hydroxide ); an adhesive entity such as, for example, a muco phase of polymer matrix is prepared by adding the appro or bio - adhesive ; excipients to maximize solubility of active priate quantities of sorbitol ( and surfactant and / or active pharmaceutical compound ( s ) ; an antigen ( s ) and /or an adju agent, if desired ) to water , heating to approximately 60 -75° vant( s ) to induce an intestinal mucosal or a systemic C . until in solution and then adding gelatin although the immune response . precise order and timing of addition is not critical. A typical [0340 ] The composition may further comprise excipients “ 8gelatin solution ” comprises 15 -25 % (preferably 17 - 18 % ) to enhance the therapeutic potential of active agents in the gelatin ; 75 % -85 % (preferably 77 - 82 % ) of water plus from ileum and colon including , but not limited to absorption 1 - 5 % (preferably 1. 5 to 3 % ) sorbitol. US 2018 / 0042989 A1 Feb . 15 , 2018
[0346 ] The choice of temperature at which the colloid is ( e . g . Solutol or Cremophor RH40 ) in the dispersed phase , formed depends however on various factors include the slight warming e . g . to 40 - 50° C . is appropriate. temperature lability of the active pharmaceutical ingredient [0350 ] The colloid may be formed by addition of the and the amount of plasticiser included in the gelatin , the type dispersed phase to the heated aqueous phase with stirring as of gelatin , as well as other factors . Generally however, the described above . The resultant colloid then has the compo gelatin solution ( especially in the case of standard or normal sition of the solidified minibeads described above but with gelatin ) is maintained at 60° C . -70° C . to maintain it in a water still present. fluid state . [0351 ] The colloid is then poured or introduced into a [0347 ] The processing temperature can be reduced to a mould or other vessel or poured onto sheets or between desirable target temperature e . g . 37° C . by use of lower sheets or delivered dropwise (or extruded ) into another fluid melting -point gelatin (or gelatin derivatives or mixtures of such that the polymer matrix - containing aqueous phase , on gelatins with melting point reducers ) or other polymer solidification , takes the form of the mould , vessel, sheet or matrix material such as, for example , sodium alginate for droplet/ bead intended . It is preferred to progress to mould example when the active principle to be incorporated in the forming e . g . beading without delay . composition of the invention is temperature -labile . Alterna [0352 ] Alternatively to moulding , specialised machinery tively, temperature - labile active principlesmay be processed can be employed for example to create the hemispherical at higher temperatures by using appropriate apparatus or beads described above (see section above entitled " Shape , machinery which limits the time during which the tempera Size and Geometry ” ) in which the invention takes the form ture - labile active principle is in contact with the higher ofhemispherical beads . It is possible to manufacture a single temperature medium . For example , if gelatin droplets are bead made from joining two such hemispheres (i . e . a single being formed by machine extrusion and immediately cooled bead having two distinct halves ) by using specialist appa e . g . in a cooling bath , additional appropriate inlet tubing can ratus in which two tubes through which two different emul be used to introduce temperature - sensitive active principle sions are flowing , normally of circular cross section , are into the fluid gelatin solution ( and the mixture can be joined shortly before an extrusion point or nozzle ( which immediately homogenized ) very shortly before ejection may be vibrating ) into a single dual lumen tube with a flat from a beading nozzle or other dropletting process such that wall separating the two emulsion flows and which prevents the duration of exposure of the active principle to the higher the two emulsions from coming into contact until the point temperature gelatin is limited so reducing the degree of any of extrusion . The cross - section of the joined dual- lumen tube up to the point of extrusion therefore appears as two semi heat - dependent degradation of the active principle . This circles . In operation , the two hemispherical emulsion flows process may use any appropriate device such as , for combine to form a single , substantially spherical , bead on example , a homogenizer , e .g . a screw homogenizer, in extrusion such that normal droplets are ejected / extruded for conjunction with an extrusion - type apparatus as described solidification . for example in WO 2008 / 132707 (Sigmoid Pharma ) the [0353 ] Solidification can occur in a variety of ways entirety of which is incorporated herein by reference . depending on the polymer of the matrix , for example by [ 0348 ] Hydrophobic surfactant, if included , is added to the changing the temperature around the mould , vessel, sheet, aqueous phase conveniently at the same time the other droplet/ bead etc or by applying a solidification fluid or components are added e . g . polymer matrix material and hardening solution so that the moulded shape is gelled or plasticiser if included e . g . at the beginning of the processing solidified . In certain embodiments both temperature change session . The physical form of the surfactant at the point of and application of a solidifying fluid or hardening solution introduction into the aqueous phase during preparation may are employed together or simultaneously . play a role in the ease of manufacture of the composition [0354 ] In the preferred embodiment in which the compo according to the invention . As such , although liquid surfac sition of the invention takes the form of minibeads , the tants can be employed , it is preferred to utilize a surfactant minibeads may be formed for example by dropping the which is in solid form (e .g . crystalline or powder) at room liquid colloid dropwise into a fluid which effects solidifica temperature , particularly when the aqueous phase comprises tion . Where the viscosity of the colloid to be beaded reaches gelatin . Surfactant is added in the appropriate amount a certain point, drop formation becomes more difficult and required to achieve the proportion desired and as described specialised apparatus is then preferred . above . In general this leads to presence of surfactant in an 10355 ] In the case where solidification can be achieved by amount between 0 .8 % and 1 % (by weight) of the aqueous raising or reducing temperature , the temperature of the phase . solidification fluid can be adapted to achieve solidification at [ 0349 ] The dispersed phase material need not be heated the desired rate . For example , when gelatin is used as the unless it is ( semi- ) solid at ambient temperature and any polymer matrix , the solidification fluid is at a lower tem active principle and in this case other dispersed phase perature than the temperature of the emulsion thus causing components are usually added at ambient temperature with solidification of the polymer matrix . In this case , the solidi stirring until clear. These other components may include a fication fluid is termed a cooling fluid . volatile (or non - volatile ) solvent in addition to the solvent (0356 ] In the case where solidification can be achieved and / or surfactant if selected . The appropriate amount of oil chemically , e . g . by induction of cross - linking on exposure to phase active principle ( if any ) is added to achieve the target a component of the solidification fluid , the concentration of proportion . Stirring can continue for a few minutes to a few such component in the solidification fluid and /or its tem hours , even overnight, depending on the active principle (for perature (or other characteristic or content ) can be adjusted example , an active may take several hours to be fully to achieve the desired rate and degree of solidification . For dissolved ) . Where it is desired to include an oil e . g . a wax example , if alginate is chosen as the polymer matrix , one oil which is not liquid or fully liquid at room temperature component of the solidification fluid may be a calcium US 2018 / 0042989 A1 Feb . 15 , 2018 27 containing entity ( such as , for example , calcium chloride ) achieved by the addition of an inactive form of the ion that able to induce cross- linking of the alginate and consequent will cause crosslinking of the alginate , which is then acti solidification . Alternatively , the same or similar calcium vated by a change in e . g . pH after sufficient dispersion ofthe containing entity may be included ( e . g . dispersed ) in the ion is complete (see Glicksman , 1983a ; Hoefler, 2004 which aqueous phase of the liquid colloid prior to beading and are both incorporated herein by reference ). This approach is triggered to induce cross - linking e . g . by applying a higher or particularly useful where rapid gelation is desired and / or lower pH to a solidification fluid into which droplets of where the diffusion approach may lead to loss of API by emulsion fall dropwise or are introduced . Such electrostatic diffusion thereof into the crosslinking bath . cross - linking can be varied as to the resulting characteristics [0362 ] Following shape -forming , moulding or beading , of the minibead by control of calcium ion availability the resultant shapes or forms may be washed then dried if ( concentration ) and other physical conditions (notably tem appropriate. In the case of minibeads solidified in a solidi perature ). The solidification fluid may be a gas ( for example fication fluid , an optional final step in the method of pro air ) or a liquid or both . For example , when gelatin is used as duction described above therefore comprises removal of the the polymer matrix , the solidification fluid can be initially solidified minibeads from the solidification fluid . This may gaseous (e . g . droplets passing through cooling air ) and then be achieved e . g . by collection in a mesh basket through subsequently liquid ( e . g . droplets passing into a cooling which the solidification fluid ( e . g . MCT ) is drained and the liquid ) . The reverse sequence may also be applied while beads retained and is preferably conducted without delay gaseous or liquid cooling fluids alone may also be used . e . g . as soon as the beads have formed or within 5 , 10 , 15 , 20 , Alternatively , the fluid may be spray -cooled in which the 25 or 30 minutes of their formation . Excess solidification colloid is sprayed into a cooling gas to effect solidification . fluid may then be removed using a centrifuge (or other [ 0357] In the case of gelatin or other water- soluble poly apparatus or machine adapted to remove excess fluid ) fol mer destined to form the immobilization matrix , it is pre lowed by drying of the beads to remove water or free water ferred that the solidification fluid be a non - aqueous liquid and /or removal of some or all of any additional solvent e .g . ( such as , for example , medium chain triglycerides , mineral ethanol or isopropyl alcohol used to dissolve or facilitate oil or similar preferably with low HLB to ensure minimal dissolution of the active principle in preceding steps option wetting) which can conveniently be placed in a bath (cooling ally followed by washing ( e . g . using ethyl acetate ) and a bath ) to receive the droplets of emulsion as they solidify to subsequent " drying ” step to remove excess solvent ( e . g . form beads . Use of a non - aqueous liquid allows greater ethyl acetate ). Isopropyl alcohol is an example of a solvent flexibility in choice of the temperature at which cooling is which is preferably removed later in processing to reduce conducted . residues in the oil or aqueous phase . Drying can be achieved [0358 ] Where a liquid cooling bath is employed , it is by any suitable process known in the art such as use of a generally maintained at less than 20° C ., preferably main drum drier ( e . g . Freund Drum dryer which may be part of the tained in the range 5 - 15° C . , more preferably 8 - 12° C . when Spherex equipment train if used ) with warm air at between standard gelatin is used as the polymer matrix . If a triglyc 15° C . and 25° C ., preferably around 20° C . leading to eride is chosen as the cooling fluid in the cooling bath , a evaporation or entrainment of the water by the air. Use of preferred example is Miglyol 810 from Sasol. gelatin as the polymer matrix ( e . g . as principal constituent of [0359 ] If gelatin is selected as the polymer matrix , respect the aqueous immobilisation phase ) in most cases requires a for appropriate temperature ranges ensures solidification of drying step and for minibeads this is preferably achieved by the gelatin at an appropriate rate to avoid destruction e . g . of drying in air as above described . The resultant composition tertiary protein structure in the case where the active prin (the composition of the invention ) is essentially dry as ciple is a protein . described in more detail above. [0360 ] If alginate is selected as the polymer matrix , a [ 0363 ] In terms of the way in which colloid droplets may typical method of making minibeads involves dropwise be formed in the first step of the beading process described addition of a 3 % sodium alginate solution in which oil above , variations of the above described method are possible droplets are dispersed as described above into a 4° C . including introducing droplets into a variety of solidification crosslinking bath containing 0 . 1 M calcium chloride to fluids. produce calcium alginate ( this method can be referred to as f0364 ] In general, the minibeads may be generated by the " diffusion setting ” because the calcium is believed to diffuse application of surface tension between the fluid colloid into the minibeads to effect cross - linking or setting ) . Using having an aqueous continuous phase and an appropriate a syringe pump, or Inotech machine , droplets can be gen solidification fluid such as , for example , gas or liquid in erated or extruded ( e . g . at 5 mL / h if a pump is used ) through order to create the spherical or substantially spherical shape a sterile needle or other nozzle (described elsewhere herein ) of the ultimate beads. which can be vibrating as discussed elsewhere herein . Air [0365 ] Alternatively , the minibeads may be produced flow of between 15 and 20 L /min through 4 .5 mm tubing can through ejection or extrusion of the liquid colloid through an be applied downwards over the needle to reduce droplet size orifice or nozzle with a certain diameter and optionally if desired . Newly formed minibeads can then be stirred in subject to selected vibrational frequencies and/ or gravita the calcium chloride bath for up to an hour. If carrageenan tional flow . Examples of machines which may be used are is used as the polymer matrix both salt and reduction in the Freund Spherex , ITAS/ Lambo , Globex or Inotech pro temperature e . g . by dropping into cooling oil may be used to cessing equipment. Operation of the Spherex machine obtain solidification . manufactured by Freund as may be desired to manufacture [ 0361] An alternative approach when using alginate is minibeads according to the present invention is described in internal gelation in which the calcium ions are dispersed in U . S . Pat . No . 5 ,882 ,680 (Freund ) , the entire contents of the aqueous phase prior to their activation in order to cause which are incorporated herein by reference . It is preferred to gelation of hydrocolloid particles. For example , this can be select a vibrational frequency in the region of 10 - 15 RPM US 2018 / 0042989 A1 Feb . 15 , 2018 28 although the ultimate choice (and separately the amplitude ing conditions may be varied for example by altering the of vibration selected ) depends on the viscosity of the colloid temperature of the liquid colloid , the solidification fluid and to be beaded . If the polymer matrix is chosen to solidify at the concentration of components in these fluids and the time lower temperature , it may be appropriate to maintain the allowed for certain processing steps to occur including lines to the orifice/ nozzle at a certain temperature to main drying. Although not currently preferred , such variations tain the fluidity of the solution . may be applied in the case of minibead manufacture to 10366 ). The Spherex machine ( and others ) may be adapted achieve heterogeneity such as , for example, a harder skin to make use of a dual concentric lumen nozzle to ensure and softer core with less than complete immobilization of oil simultaneous extrusion of two fluids , the fluid in the inner droplets towards the core as opposed to the surface of the lumen forming a core and the fluid of the outer lumen bead . Larger ( e . g . non -beaded ) forms or shapes of the forming a capsule . The fluid forming the capsule is solidified composition according to the invention may particularly be according to one of the methods described . It may or may engineered to embody such heterogeneity . However , it is not be desirable for the fluid forming the core to be suscep currently preferred to have internally homogenous compo tible of solidification to yield a particular embodiment of the sitions and , within the minibead embodiment, this can be composition of the invention . The machinery adapted in this favoured by conducting the beading / dropletting using a way can be used to manufacture the composition of the homogeneous medium e . g . a well dispersed colloid . Such invention in the form of a capsule in which the core of the homogeneity in the emulsion to be beaded can help avoid the composition is filled with a fluid ( a gas or a liquid ) as drying conditions affecting symmetry . described in the section above entitled " Shape , Size and [ 0372 ] The oral composition may be used for a number of Geometry ” ( noting that the core , like the capsular material, applications as discussed elsewhere herein . The active prin may be a composition , albeit optionally a distinct compo ciple ( s ) may be released immediately immediate release sition , according to the invention i. e. susceptible of solidi profile ) or be released after some delay and/ or over an fication according to one of the methods described above ) . extended period (delayed and / or extended release profile ) . A three - lumen nozzle and appropriate tubing may be For immediate release , the minibeads or other formats may employed if it is desired to include an intermediate internal be uncoated or coated enterically to protect against stomach layer e . g . internal film layer of non - aqueous material on the acid for immediate release in the small intestine . inner face of the sphere with the intermediate layer conve [ 0373 ] Alternatively , if controlled release is desired ( i. e . niently being solid at room temperature . Thus , in terms of delayed , extended or site -targeted release etc ) , or ifmedium the softness / hardness of successive layers , the composition independent release is desired , it is possible , according to the may for example be described as solid : solid in the case of invention to apply a coat to the minibeads or other formats . two layers or solid : solid : solid in the case of 3 layers or Application of the appropriate coat may, for example if liquid / semi- liquid :solid :solid in the case of 3 layers . colonic release is required , allow for say less than 10 % of the [0367 ] The preceding paragraphs describe the formation active principle to be dissolved ( in dissolution medium ) at 4 of uncoated beads . It is a preferred embodiment of the hours and then a burst ( sudden release ) towards a maximum present invention to have coated beads which are described dissolution ( approaching 100 % ) in the subsequent 24 hours . in more detail elsewhere herein . Such coatings may be single Many alternative target profiles are possible and this or multiple and may be applied in a number of ways ( see example is purely for illustration . separate section ) . [03741 . Thus , the composition may be in the form of [0368 ] With regard to one of the methods described above minibeads at least some of which bear a coat (i . e . are coated ) ( ejection of colloid through an optionally vibrating nozzle ) in order to control release of active principle from the with two concentric orifices ( centre and outer ), the outer minibead . In one embodiment, the coat is a film and in fluid may form a coat (outside the minibead ) of e .g . poly another embodiment, it is a membrane . The coat, film or meric material ( polymeric coating ) which may contain an membrane comprises one or more substances preferably of active principle or may impart controlled release character a polymeric nature ( e . g . methacrylates etc; polysaccharides istics to the minibead and the inner layer (core ) may be a etc as described in more detail below ) or combination of colloid as described herein . The Spherex machine manufac more than one such substance , optionally including other tured by Freund ( see U . S . Pat . No. 5 , 882 ,680 to Freund ) is excipients or active principles , such as , for example , plas preferably used ( the entire contents of this patent is incor ticizers , described e . g . in the sections above on active porated herein by reference ). principles. Preferred plasticizers, if they are used , include [0369 ] Use of the Spherex machine achieves very high hydrophilic plasticizers for example triethyl citrate ( TEC ) monodispersity . For example , in a typical 100 g , batch 97 g which is particularly preferred when using the Eudragit ofminibeads were between 1. 4 to 2 mm diameter or between family of polymers as coatings as described below . Another 1 and 2 mm . Desired size ranges can be achieved by methods preferred plasticiser, described in more detail below in known in the art for rejecting/ screening different sized relation to coating with ethyl cellulose, is DBS . Alternative particles . For example , it is possible to reject/ screen out the or additional optionally included excipients are glidants . A larger /smaller beads by passing a batch first through e . g . a 2 glidant is a substance that is added to a powder or other mm mesh and subsequently through a 1 . 4 mm mesh . medium to improve its flowability . A typical glidant is talc [ 0370 ] The 1 . 4 to 2 mm diameter range is a good size if it which is preferred when using the Eudragit family of is desired to coat the minibeads ( if smaller , the spray of the polymers as coatings . coating machine may bypass the minibead ; if too large hard , the beads may be harder to fluidise which is necessary to achieve consistent coating ). Non -Colloidal Formulations [0371 ] The minibeads are preferably internally ( i . e . cross [0375 ] In the case of non -colloidal formulations compris sectionally ) homogeneous i. e . monolithic although process ing a water - soluble polymer matrix , the samemethods may US 2018 / 0042989 A1 Feb . 15 , 2018 29 be followed as just described for colloidal formulations but fluids independently of pH . In the swollen state , they are without the involvement of a dispersed phase. then permeable to water and dissolved active agents . The permeability of the polymers depends on the ratio of ethy Coating lacrylate (EA ) , methyl methacrylate (MMA ), and trimeth [0376 ] In the case of combinations of polymers, combi ylammonioethyl methacrylate chloride ( TAMCI) groups in nations may be selected in order to achieve the desired delay the polymer . For example , those polymers having EA : M (or other change ) in the release of the drug and / or poration MA: TAMC1 ratios of 1 : 2 : 0 . 2 ( EUDRAGITTM RL ) are more of the coating and / or exposure of the minibead or other permeable than those with ratios of 1 : 2 : 0 . 1 (EUDRAGITTM format within the coating to allow egress of drug and/ or RS ). Polymers of EUDRAGITTM RL are insoluble polymers dissolution of the immobilization matrix . In one embodi of high permeability . Polymers of EUDRAGITTM RS are ment, two types of polymers are combined into the same insoluble filmsof low permeability . A particularly preferred polymeric material, or provided as separate coats that are diffusion - controlled pH - independent polymer in this family applied to the minibeads . is RS 30 D which is a copolymer of ethyl acrylate, methyl [0377 ] It has previously been stated that the formulations methacrylate and a low content of methacrylic acid ester may comprise more than one population of minibeads. with quaternary ammonium groups present as salts to make Within the coating embodiment, the differences between the polymer permeable . RS 30 D is available as an aqueous populations may lie in the coat i. e . two (or more ) populations dispersion . of minibeads may differ in a number of respects one of which is the coating . [ 0383 ] The amino methacrylate co -polymers can be com [ 0378 ] The coat may be applied as described below and bined in any desired ratio , and the ratio can be modified to may vary as to thickness and density . The amount of coat is modify the rate of drug release . For example , a ratio of defined by the additional weight added to ( gained by ) the EUDRAGITTM RS : EUDRAGITTM RL of 90 : 10 can be used . dried composition ( e . g . minibead ) of the invention . Weight Alternatively, the ratio of EUDRAGITTM gain is preferably in the range 0 . 1 % to 50 % , preferably from RS: EUDRAGITTM RL can be about 100 : 0 to about 80 : 20 , or 1 % to 15 % of the dry weight of the bead , more preferably about 100 : 0 to about 90 : 10 , or any ratio in between . In such in the range 3 % to 10 % or in the range 5 - 12 % or in the range formulations , the less permeable polymer EUDRAGITTM 8 - 12 % . RS generally comprises the majority of the polymeric mate [ 0379 ] The polymeric coating material may comprise rial with the more soluble RL , when it dissolves, permitting methacrylic acid co - polymers , ammonio methacrylate co gaps to be formed through which solutes can come into polymers , or mixtures thereof. Methacrylic acid co -poly contact with the minibead allowing pre - dissolved pharma mers such as , for example , EUDRAGITTM S and ceutical actives to escape in a controlled manner. EUDRAGITTM L (Evonik ) are particularly suitable . These [ 0384 ] The amino methacrylate co -polymers can be com polymers are gastroresistant and enterosoluble polymers . bined with the methacrylic acid co - polymers within the Their polymer films are insoluble in pure water and diluted polymeric material in order to achieve the desired delay in acids . They may dissolve at higher pHs, depending on their the release of the drug and / or poration of the coating and / or content of carboxylic acid . EUDRAGITTM S and exposure of the minibead within the coating to allow egress EUDRAGITTM L can be used as single components in the of drug and /or dissolution of the immobilization or water polymer coating or in combination in any ratio . By using a soluble polymer matrix . Ratios of ammonio methacrylate combination of the polymers , the polymeric material can co - polymer ( e . g . , EUDRAGITTM RS) to methacrylic acid exhibit solubility at a variety of pH levels , e . g . between the co - polymer in the range of about 99: 1 to about 20: 80 can be pHs at which EUDRAGITTM L and EUDRAGITTM S are used . The two types of polymers can also be combined into separately soluble . the same polymeric material, or provided as separate coats [0380 ] The trademark “ EUDRAGIT” is used hereinafter that are applied to the minibeads. to refer to methacrylic acid copolymers , in particular those sold under the EUDRAGITTM by Evonik . [0385 ] EudragitTM FS 30 D is an anionic aqueous- based [ 0381 ] The coating can comprise a polymeric material acrylic polymeric dispersion consisting of methacrylic acid , comprising a major proportion ( e . g . , greater than 50 % of the methyl acrylate, and methyl methacrylate and is pH sensi total polymeric coating content) of at least one pharmaceu tive . This polymer contains fewer carboxyl groups and thus tically acceptable water - soluble polymer , and optionally a dissolves at a higher pH ( > 6 . 5 ) . The advantage of such a minor proportion ( e . g ., less than 50 % of the total polymeric system is that it can be easily manufactured on a large scale content) of at least one pharmaceutically acceptable water in a reasonable processing time using conventional powder insoluble polymer. Alternatively , the membrane coating can layering and fluidized bed coating techniques . A further comprise a polymeric material comprising a major propor example is EUDRAGIT® L 30D -55 which is an aqueous tion ( e . g . , greater than 50 % of the total polymeric content ) dispersion of anionic polymers with methacrylic acid as a of at least one pharmaceutically acceptable water insoluble functional group . It is available as a 30 % aqueous disper polymer, and optionally a minor proportion ( e . g . , less than sion . 50 % of the total polymeric content) of at least one pharma 0386 ). In addition to the EUDRAGITTM polymers ceutically acceptable water - soluble polymer. described above , a number of other such copolymers can be [ 0382 ] Ammonio methacrylate co - polymers such as , for used to control drug release . These include methacrylate example , EUDRAGITTM RS and EUDRAGITTM RL ester co - polymers such as , for example , the EUDRAGITTM ( Evonik ) are suitable for use in the present invention . These NE and EUDRAGITTM NM ranges . Further information on polymers are insoluble in pure water, dilute acids , buffer the EUDRAGITTM polymers can be found in “ Chemistry solutions , and / or digestive fluids over the entire physiologi and Application Properties of Polymethacrylate Coating cal pH range . The polymers swell in water and digestive Systems ," in Aqueous Polymeric Coatings for Pharmaceu US 2018 / 0042989 A1 Feb . 15 , 2018 30 tical Dosage Forms, ed . James McGinity, Marcel Dekker order to modify the dissolution of the coated minibead . Inc . , New York , pg 109 - 114 the entirety of which is incor Unless otherwise stipulated , use of the term “ Surelease ” porated herein by reference . may apply to Surelease E - 7 - 19020 , E - 7 - 19030 , E - 7 - 19040 [0387 ] Several derivatives of hydroxypropyl methylcellu or E - 7 - 19050 . E - 7 - 19020 comprises ethylcellulose blended lose (HPMC ) also exhibit pH dependent solubility and may with oleic acid and dibutyl sebacate , then extruded and be used in the invention for coating. These include hydroxy melted . The molten plasticized ethylcellulose is then directly propyl methylcellulose phthalate (HPMCP ) , which rapidly emulsified in ammoniated water in a high shear mixing dissolves in the upper intestinal tract and hydroxypropyl device under pressure . Ammonium oleate is formed in situ methylcellulose acetate succinate (HPMCAS ) in which the to stabilize and form the dispersion of plasticized ethylcel presence of ionizable carboxyl groups causes the polymer to lulose particles. Additional purified water is then added to solubilize at high pH ( > 5 . 5 for the LF grade and > 6 . 8 for the achieve the final solids content. E - 7 - 19030 additionally HF grade ). These polymers are commercially available from comprises colloidal anhydrous silica dispersed into the Shin - Etsu Chemical Co . Ltd . As with other polymers material. E -7 - 19040 is like E - 7 - 19020 except that it com described herein as useful for coatings, HPMC and deriva prises medium chain triglycerides instead of dibutyl seba tives may be combined with other polymers e . g . cate . E - 7 - 19050 derives from blending ethylcellulose with EUDRAGIT RL -30 D . oleic acid before melting and extrusion . The molten plasti [0388 ] It is particularly preferred according to the inven cized ethylcellulose is then directly emulsified in ammoni tion to use a polymeric coating substance which is pH ated water in a high shear mixing device under pressure . independent in its dissolution profile and / or in its ability to Ammonium oleate is formed in situ to stabilize and form the release active principles incorporated in the minibeads of the dispersion of plasticized ethylcellulose particles . However, invention . Examples have already been given ( e . g . , Eudragit E - 7 - 19040 is preferred . RS and RL ) . Another example of a pH - independent poly [ 0391 ] The invention also contemplates using combina meric coating substance is ethylcellulose , in particular a tions of Surelease with other coating components, for dispersion of ethylcellulose in a sub -micron to micron example sodium alginate , e . g . sodium alginate available particle size range , e . g . from about 0 .1 to 10 microns in size , under the trade name NutratericTM . homogeneously suspended in water with the aid of an [0392 ] In addition to the EUDRAGITTM and Surelease® emulsification agent, e . g . ammonium oleate . The ethylcel polymers discussed above , other enteric , or pH - dependent, lulose dispersion may optionally and preferably contain a polymers can be used . Such polymers can include phthalate , plasticizer , for example dibutyl sebacate (DBS ) or medium butyrate, succinate, and / or mellitate groups. Such polymers chain triglycerides . Such ethylcellulose dispersions may , for include , but are not limited to , cellulose acetate phthalate , example , be manufactured according to U . S . Pat. No . 4 ,502 , cellulose acetate succinate , cellulose hydrogen phthalate , 888 , which is incorporated herein by reference . One such cellulose acetate trimellitate , hydroxypropyl- methylcellu ethylcellulose dispersion suitable for use in the present lose phthalate , hydroxypropylmethylcellulose acetate succi invention and available commercially is marketed under the nate , starch acetate phthalate , amylose acetate phthalate , trademark Surelease , by Colorcon of West Point, Pa . USA . polyvinyl acetate phthalate , and polyvinyl butyrate phtha In this marketed product , the ethylcellulose particles are , late . Additionally , where compatible , any combination of e . g . , blended with oleic acid and a plasticizer, then option polymer may be blended to provide additional controlled - or ally extruded and melted . The molten plasticized ethylcel targeted - release profiles . lulose is then directly emulsified , for example in ammoni 10393 ] The coating can further comprise at least one ated water optionally in a high shear mixing device , e . g . soluble excipient to increase the permeability of the poly under pressure . Ammonium oleate can be formed in situ , for meric material . Suitably , the at least one soluble excipient is instance to stabilize and form the dispersion of plasticized selected from among a soluble polymer , a surfactant, an ethylcellulose particles . Additional purified water can then alkali metal salt , an organic acid , a sugar, and a sugar be added to achieve the final solids content. See also U . S . alcohol. Such soluble excipients include , but are not limited Pat. No . 4 , 123 ,403 , which is incorporated herein by refer to , polyvinyl pyrrolidone , polyethylene glycol, sodium chlo ence . ride , surfactants such as, for example , sodium lauryl sulfate [0389 ] The trademark “ Surelease® " is used hereinafter to and polysorbates , organic acids such as, for example , acetic refer to ethylcellulose coating materials , for example a acid , adipic acid , citric acid , fumaric acid , glutaric acid , dispersion of ethylcellulose in a sub -micron to micron malic acid , succinic acid , and tartaric acid , sugars such as , particle size range , e . g . from about 0 . 1 to 10 microns in size , for example , dextrose , fructose , glucose , lactose , and homogeneously suspended in water with the aid of an sucrose , sugar alcohols such as , for example , lactitol, malti emulsification agent, e . g . ammonium oleate . In particular , tol, mannitol , sorbitol , and xylitol , xanthan gum , dextrins, the trademark “ Surelease® " is used herein to refer to the and maltodextrins. In some embodiments , polyvinyl pyrroli product marketed by Colorcon under the Surelease® trade done, mannitol, and /or polyethylene glycol can be used as mark . soluble excipients . The at least one soluble excipient can be [0390 ] Surelease® dispersion is an example of a combi used in an amount ranging from about 1 % to about 10 % by nation of film - forming polymer, plasticizer and stabilizers weight, based on the total dry weight of the polymer. which may be used as a coating to adjust rates of active [0394 ] The modifications in the rates of release, such as to principle release with reproducible profiles that are rela create a delay or extension in release , can be achieved in any tively insensitive to pH . The principal means of drug release number of ways. Mechanisms can be dependent or indepen is by diffusion through the Surelease dispersion membrane dent of local pH in the intestine , and can also rely on local and is directly controlled by film thickness . Use of Sure enzymatic activity to achieve the desired effect. Examples of lease® is particularly preferred and it is possible to increase modified -release formulations are known in the art and are or decrease the quantity of Surelease applied as coating in described , for example , in U .S . Pat. Nos . 3 ,845 , 770 ; 3, 916 , US 2018 / 0042989 A1 Feb . 15 , 2018 31
899 ; 3 , 536 , 809 ; 3 ,598 , 123 ; 4 ,008 ,719 ; 5 ,674 , 533 ; 5 ,059 , particularly preferred polysaccharide in this embodiment of 595 ; 5 ,591 , 767 ; 5 ,120 ,548 ; 5 ,073 ,543 ; 5, 639 , 476 ; 5, 354 , the present invention is pectin . Various kinds of pectin may 556 ; and 5 ,733 , 566 all of which are incorporated herein by be used including pectin of different grades available i. e . reference in their entirety . with differing degrees of methylation ( DM ), i. e . percentage [ 0395 ] As noted above , Surelease is a particularly pre of carbonyl groups esterified with methanol, for example ferred polymer coating owing to its pH -independent disso pectins with a DM of more than 50 % , known as High lution character. However, the inventors /applicants have Methoxy (HM ) Pectins or Low Methoxy (LM ) pectins, or a found that it is difficult to select the appropriate amount pectin combination comprising an HM pectin and an LM (weight gain ) of Surelease to achieve optimal dissolution . It pectin . It is also possible in this embodiment to use pectins has been found that too much Surelease leads to incomplete having various degrees of acetylation (DAC ). Taken ( or over slow ) dissolution while too little leads to over fast together , the DM and DAC or the degree of substitution is dissolution . known as Degree of Esterification (DE ) . Pectins of various [ 0396 ] The inventors /applicants have found that addition DE ' s may be used according to the invention . As an alter to SureleaseTM of a second polymer ( e . g . a polysaccharide , native to pectin , sodium alginate may be used as a polysac especially a heteropolysaccharide ) which is normally charide according to an embodiment of the invention . How degraded by bacterial enzymes but not by digestive ever , other embodiments may conveniently include amylose enzymes , e .g . human digestive enzymes, resolves this prob and / or starch which contains amylose . Various grades of lem and provides flexibility in modulating the amount of starch , containing different percentages of amylose may be polymer added to the minibeads of the invention in order to used including for example Hylon V (National Starch Food achieve optimal dissolution profiles . In general terms, there Innovation ) which has an amylose percentage of 56 % or fore, the disclosure includes formulations as described Hylon VII which has an amylose percentage of 70 % . The herein which comprise a coating comprising a combination remaining percentage is amylopectin . The polysaccharides of a delayed release material, for example an erodible pectin , amylose and sodium alginate are particularly pre polymer e . g. ethylcellulose , and a polymer susceptible of ferred for achieving colon delivery i . e . for compositions degradation by bacterial enzymes in the colon , e . g . a poly intended to release active principles in the colon . saccharide and particularly a water soluble polysaccharide , [0399 ]. It has been found that pectin can act as a former of particularly a pectin . pores in the coating otherwise provided by ethylcellulose [0397 ] The disclosure therefore includes a coating for (preferably Surelease ) . By “ pores” is not meant shaft - like compositions intended to release their active payload in the holes from the surface to the core of the minibead , rather colon which is a combination of ethylcellulose (preferably areas of weakness or absence of coating occurring stochas formulated with an emulsification agent such as, for tically on and within the coating of the invention . example , ammonium oleate and /or a plasticizer such as, for [0400 ) Pore formers have been described before in con example , dibutyl sebacate or medium chain triglycerides ) nection with Surelease ( see e . g . US 2005 /0220878 ) but in and a polysaccharide susceptible of degradation by a bac relation to " gastro - insoluble ” substances such as , for terial enzyme normally found in the colon . Such polysac example , alginate . charides include chondroitin sulphate , pectin , dextran , guar [ 0401 ] Where the water - soluble polysaccharide (WSP ) is gum and amylase , chitosan etc and derivatives of any of the pectin , the proportion of ethylcellulose or SureleaseTM to foregoing. Chitosan is particularly preferred in connection pectin is ideally in the range 90 : 10 to 99 : 1 , preferably , 95 :5 with obtaining a colon - specific release profile . The disclo to 99 : 1 , more preferably 98 : 2 to 99 : 1 . sure also includes a composition comprising a combination [0402 ] In this particularly preferred combination ( ethyl of ethylcellulose ( preferably formulated with an emulsifica cellulose or SureleaseTM + WSP e . g . pectin ) the weight gain tion agent such as , for example , ammonium oleate and /or a and ratio between ethylcellulose or SureleaseTM and WSP plasticizer such as , for example , dibutyl sebacate or medium can be varied to refine the behaviour of the coating and the chain triglycerides ) and a polysaccharide susceptible of composition of the invention when it bears such a coat. Thus degradation by a bacterial enzyme normally found in the to the inventors / applicant' s surprise , the advantages of this colon ; the composition may include a liquid vehicle , e . g . preferred combination of coating polymers were further water. pronounced by selecting a weight gain in the range 0 to 30 % [0398 ] The use of polysaccharides by themselves for coat (preferably 5 to 10 % ) and a weight ratio of ethylcellulose or ing purposes has been tried with limited success . Most of the Surelease to pectin in the range 95 : 5 to 99. 5 :0 .5 preferably non -starch polysaccharides suffer from the drawback of 97 : 3 to 99 :1 inclusive. Particularly favoured weight gains lacking good film forming properties . Also , they tend to using ethylcellulose or Surelease are those in the range swell in the GI tract and become porous, resulting in the 5 - 12 % or in the range 8 - 12 % . early release of the drug . Even amorphous amylose, which [0403 ] Although the focus above has been on extending is resistant to degradation by pancreatic alpha amylase but and /or sustaining release of active principles from minibeads capable of degradation by colonic bacterial enzymes has the or other formats , also contemplated are uncoated or simple disadvantage of swelling in aqueous media although this can enteric coated minibeads or other formats providing early , be controlled by incorporating insoluble polymers like , ethyl small intestinal API release with sufficient enteric coating cellulose and acrylates into the amylose film . Amylose merely to protect the minibeads from dissolution in the however is not water - soluble and although water -soluble stomach . polysaccharides are not excluded , the present inventors have [0404 ] It is preferred to dry the minibeads before they are found that use of a water - soluble polysaccharide (WSP ) coated with a suitable polymeric coat (as described in more susceptible of bacterial enzymic degradation brings particu detail above /below ) . It is also preferred , in certain embodi larly advantageous results when used as a coating in accor ments to apply a first coat before applying a second . In dance with this embodiment of the present invention . A general the first coat and the second coat may be of the same US 2018 / 0042989 A1 Feb . 15 , 2018 32 or differentmaterials and be chosen from any of the classes - continued of coating material described herein . In specific embodi ments , the first coat optionally protects the core (bead ) from Process Parameter Values interaction with the second coat and/ or prevents leaching of Exhaust air temperature ( ° C .) 38 - 42 bead contents into the second coat. For example , the first Product temperature (°C . ) 38 - 42 coat may comprise or consist of a mixture of hypromellose , Atomizing air pressure (bar ) Up to 1 . 4 e . g . 0 . 8 - 1 . 2 titanium dioxide and polyethylene glycol; the second ( outer ) Spray rate ( g /min ) 2 - 10 and 3 - 25 RPM coat in this instance may comprise , or consist of , a mixture of ethylcellulose and pectin or another polymer, e. g . water [0408 ] Whether as part of the polymeric coat or indepen soluble polysaccharide, susceptible to degradation by bac dently thereof, the minibeads of the invention may be coated terial enzymes in the colon , and in particular may be the with additional drug layers using methods conventional in Surelease® - pectin mixture described above . If it is desired the art of pharmaceutical science ( such as for example using for the first coat to use a mixture of hypromellose , titanium coating machines as just described ) to produce a composi dioxide and polyethylene glycol, commercial products cor tion having one or more layer ( s ) , each layer containing one responding to such mixtures are available including or more active pharmaceutical or other ingredient/ excipient Opadry® White , a product commercialised by Colorcon . as described elsewhere herein . Drug layering means the More generally , various products commercialised under the deposition of at least one or successive layers of drug entities trade name Opadry and Opadry II . Further nonlimiting from solution , suspension or dry powder on nuclei e . g . examples include Opadry YS - 1 -7706 - G white , Opadry Yel minibeads as described herein . Drug layering includes solu low 03692357 , Opadry Blue 03690842 ) . These composi tion /suspension layering , powder layering and powder drug tions are available as dry film coating compositions that can layering . In solution / suspension layering , drug particles are be diluted in water shortly before use . Opadry and Opadry dissolved or suspended in a binding liquid . In powder II formulations comprise a cellulosic film forming polymer layering , complete dissolution does not occur , due to low ( e . g . , HPMC and / or HPC ) , and may contain polydextrose , liquid saturation , irrespective of the solubility of the active maltodextrin , a plasticizer ( e . g . , triacetin , polyethylene gly agent in the binding liquid . In powder drug layering , a binder col) , polysorbate 80 , a colorant ( e . g . , titanium dioxide , one solution is first sprayed onto previously prepared inert seeds or more dyes or lakes) , and /or other suitable film - forming e . g . minibeads as described herein , followed by the addition polymers ( e . g ., acrylate -methacrylate copolymers ) . Suitable of powder. Conventional pan coaters may be used as OPADRY or OPADRY II formulations may comprise a described above for polymer coating although modified plasticizer and one or more of maltodextrin , and polydex forms of pan coaters are preferred including fluidised -bed trose ( including but not limited to a ) triacetin and polydex and centrifugal rotary granulators . Examples of suitable trose or maltodextrin or lactose , or b ) polyethylene glycol granulators include the Rotor granulator. (Glatt ) , the Rotor and polydextrose or maltodextrin ) . Particularly preferred processor ( Aeromatic ) , the Spir - a - Flow (Freund ) and the commercial products are Opadry White (HPMC /HPC CF - granulator (Freund ) . In applying a drug layer , the drug to based ) and Opadry II White ( PVA /PEG -based ) . Alternative be layered onto the minibead may optionally first be ( non - Opadry ) products for initial protective coats include admixed with appropriate excipients such as , for example , polyvinyl alcohol- polyethylene glycol graft copolymers binders as described elsewhere herein . A particularly pre such as is available commercially under the name Kollicoat ferred binder in this context is polyvinyl pyrrolidone ( also IR and methyl methacrylate ammonium -based copolymers spelt polyvinylpyrrolidone and also known as PVP or povi such as are available commercially under the name Eudragit done ) . PVPs of various K - values may be used . The K - value E . Another preferred example is low molecular weight of PVP is a function of its average molecular weight, the HPMC . The optional inner coat is applied in the same degree of polymerization , and the intrinsic viscosity . It is manner as is the outer ( or sole ) coat ( or coating layer ) . particularly preferred to use PVP K - 32. Up to 5 % of the dry 10405 ] The coating process can be carried out by any weight of the composition of the invention in this embodi suitable means such as, for example , by use of a coating ment may be made up of such binders. Approximately 1 % machine which applies a solution of a polymer coat ( as or less is preferred . Other suitable binders which may be described above in particular ) to the minibeads. Polymers used in drug - layering include gelatin , carboxymethyl cellu for coating are either provided by the manufacturer in lose , hydroxypropyl methylcellulose and hydrolysed ready -made solutions for direct use or can be made up before starches e . g . maltodextrins . Compositions embodying drug use following manufacturers ' instructions . layering may also optionally be coated with a polymer [0406 ] Appropriate coating machines are known to per coating , or include a polymer layer , to control release as sons skilled in the art and include , for example , a perforated described more generally above including the option to pan or fluidized bed system for example the GLATT , Vector include the same or a different active principle in this ( e . g . CF 360 EX ) , ACCELACOTA , Diosna , O ' Hara and / or polymer coat. HICOATER processing equipment. Most preferred is the [0409 ] The layered bead or minibead may have a plurality MFL /01 Fluid Bed Coater (Freund ) used in the “ Bottom of layers, e .g . 2 , 3 , 4 or 5 layers , comprising an active Spray ” configuration . principle , wherein the active principle of each layer is [ 0407 ] Typical coating conditions are as follows: selected independently from the active principle of each other layer. In one embodiment, each layer comprises the same active principle as each other layer; in another embodi Process Parameter Values ment, no two layers comprise the same active principle . The Fluidising airflow (m3 / h ) 20 - 60 (preferably 30 -60 ) term “ active principle ” in this paragraph embraces both a Inlet air temperature (° C .) 20 - 65 single active entity and a combination of active entities. The layered bead or minibead may comprise one or more poly US 2018 / 0042989 A1 Feb . 15 , 2018 33 mer layers , to control release as described more generally active ingredient being for therapeutic use in combination above. Such a polymer layer may contain an active principle therapy with a hydroxylase inhibitor. and therefore constitute a drug layer as well as a release 2 . A composition of clause 1 which comprises gelatin as the control layer. Alternatively , a polymer layer may be free of water - soluble polymer . active principle. A polymer layer , whether or not it contains 3 . A composition of clause 2 in which gelatin is substantially an active principle , may be located between the core and a the only water- soluble polymer. drug layer outside the polymer layer , or between two drug 4 . A composition of any preceding clause wherein the layers , or may form an outer layer . water- soluble matrix material is selected from a hydrocol [ 0410 ] The polymer layer may be located between the core loid , a non - hydrocolloid gum and chitosan and derivatives and the active principle layer . The polymer layer may be thereof. located externally of the active principle layer. The layered 5 . A composition of any preceding clause wherein the unit bead or minibead may comprise a plurality of active prin solid is a minibead having a diameter of not more than 10 ciple layers and , additionally or alternatively, it may com mm , e . g . of not more than 5 mm , the composition optionally prise a plurality of polymer layers . In some embodiments , comprising a plurality of said minibeads. there is at least one active principle layer which comprises 6 . A composition of clause 5 wherein the minibead is a release - controlling polymer . In some embodiments , the monolithic , optionally with layers thereon . outermost layer comprises a release - controlling polymer, 7 . A composition of clause 5 or clause 6 wherein the one or which may contain an active principle or , in another imple more minibeads comprise a controlled - release polymer . mentation , be free of active principle . 8 . A composition of clause 7 wherein the controlled - release [0411 ] The optionally coated minibeads of the invention polymer is an extended release polymer or an enteric poly may be formulated directly following their manufacture in mer. the ways described above . In an alternative embodiment, it 9 . A composition of clause 7 or clause 8 in which the may be desired to impart different properties to the mini minibead has a coat which comprises the controlled release beads and / or to a final solid dosage product. One way of polymer and optionally a polymer susceptible of degradation achieving this according to the invention is through granu by bacterial enzymes . lation e . g . to improve the flow of powder mixtures of 10 . A composition of clause 7 wherein the controlled - release minibeads with other components as e. g . described above in polymer is ethylcellulose comprised in a coating on the relation to binders . Granules of intact or broken minibeads minibead and optionally in association with an emulsifica may be obtained by adding liquids ( e . g . binder or solvent tion agent, for example ammonium oleate . solutions) and effecting a granulating step as described in the 11 . A composition of clause 10 wherein the ethylcellulose is prior art . Larger quantities of granulating liquid produce a also in association with a plasticizer , e . g . dibutyl sebacate or narrower particle size range and coarser and harder granules, medium chain triglycerides. i. e . the proportion of fine granulate particles decreases . The 12 . A composition of clause 10 or clause 11 wherein the optimal quantity of liquid needed to get a given particle size coating further comprises polymer susceptible of degrada may be chosen in order to minimise batch - to - batch varia tions . According to this embodiment, wet granulation is used tion by bacterial enzymes. to improve flow , compressibility , bio -availability , homoge 13 . A composition of clause 12 wherein the polymer sus neity , electrostatic properties , and stability of the composi ceptible of degradation by bacterial enzymes is water tion of the invention presented as a solid dosage form . The soluble , preferably pectin . particle size of the granulate is determined by the quantity 14 . A composition of any preceding clause wherein the and feeding rate of granulating liquid . Wet granulation may hydrophilic surfactant has an HLB value of at least 15 , and be used to improve flow , compressibility , bio -availability , optionally of at least 18 . and homogeneity of low dose blends, electrostatic properties 15 . A composition of any preceding clause wherein the of powders , and stability of dosage forms. A wet granulation hydrophilic surfactant is an anionic surfactant. process according to this embodiment may employ low or 16 . A composition of clause 15 wherein the anionic surfac high shear mixing devices in which a low viscosity liquid tant has an HLB value of at least 30 . (preferably water) is added to a powder blend containing 17 . A composition of clause 15 , wherein the anionic surfac binder previously dry mixed with the rest of the formulation tant comprises or is an alkyl sulfate salt . including minibeads. Alternative granulation approaches 18 . A composition of clause 16 in which the alkyl sulfate salt which may be utilized include high - shear , extrusion and is sodium dodecyl sulfate (SDS ) conventional wet granulation . 19 . A composition of any of clauses 15 to 18 in which the [0412 ] The invention further includes the subject matter of water - soluble polymer matrix material further contains a the following clauses : non - ionic surfactant having an HLB value of at least 10 but 1 . A pharmaceutical composition for oral administration , less than that of the hydrophilic surfactant. comprising a unit solid which comprises a water -soluble 20 . A composition of clause 19 wherein the non - ionic polymer matrix material in which matrix material are dis surfactant comprises a poly (oxyethylene ) group . persed a water- insoluble active ingredient selected from 21. A composition of clause 20 wherein the non - ionic calcineurin inhibitors , m Tor inhibitors and macrolide immu surfactant comprises a glycerol polyethylene glycol ricino nosuppressants ; droplets of water- immiscible liquid in leate . which the water - insoluble active ingredient is soluble ; and a 22 . A composition of any preceding clause in which the hydrophilic surfactant having an HLB value of at least 10 , water- immiscible liquid comprises a liquid lipid and a the composition being adapted to release water - insoluble solvent miscible therewith , in which solvent the water active ingredient in at least the colon and the water- insoluble insoluble active ingredient is soluble . US 2018 / 0042989 A1 Feb . 15 , 2018 34
23 . A composition of clause 22 when dependent on any of which matrix material are a water- insoluble active ingredi clauses 19 to 21 , in which the ratio of liquid lipid to ent selected from calcineurin inhibitors , m Tor inhibitors and non - ionic surfactant is in the range 1 -4 : 1 by weight, option macrolide immunosuppressants ; droplets of a water- immis ally 1 .2 - 3 .0 :1 by weight. cible liquid in which the water - insoluble active ingredient is 24 . A composition of clause 22 or clause 23 in which the soluble , and a hydrophilic surfactant having an HLB value liquid lipid is a medium chain triglyceride composition , the of at least 10 . medium chain triglyceride ( s ) being one or more triglycer 40 . A pharmaceutical composition for oral administration , ides of at least one fatty acid selected from Co - C , 2 fatty comprising a water - soluble polymer matrix material in acids . which matrix material are dispersed a water- insoluble active 25 . A composition of clause 24 wherein the liquid lipid is a ingredient selected from calcineurin inhibitors , m Tor inhibi caprylic /capric triglyceride. tors and macrolide immunosuppressants ; droplets of a 26 . A composition of clauses 22 to 25 in which said solvent water- immiscible liquid in which the water - insoluble active is miscible with both the liquid lipid and with water. ingredient is soluble , wherein the composition is adapted to 27 . A composition of clause 26 in which the solvent is release the water- insoluble active ingredient in at least the 2 - ( 2 - ethoxy ) ethanol. colon and is for use in treating a disorder of, or at least 28 . A composition of any preceding clause in which the suspected of being associated with , a leaky intestinal epi water - immiscible phase (water - immiscible droplets ) repre thelial barrier. sent from 10 - 85 % by dry weight of the composition . 41. A composition of clause 40 wherein the disorder is not 29 . A composition of any preceding clause in which the unit inflammatory bowel disease , Crohn ' s disease , ulcerative solid or minibead has a low water content. colitis , GVHD , or GI- GVHD . 30 . A composition of any preceding clause wherein the 42. A composition of clause 40 or clause 41 wherein the hydroxylase inhibitor is comprised in a pharmaceutical disorder is selected from celiac disease , a rheumatic disor formulation adapted to release the inhibitor in at least the der , rheumatoid arthritis , temporomandibular joint syn colon . drome, type I diabetes , multiple sclerosis , atopic dermatitis , 31 . A composition of clause 30 wherein the unit solid further psoriasis , a chronic pain syndrome, fibromyalgia , chronic comprises the hydroxylase inhibitor. fatigue syndrome, depressive disorders, affective disorders 32 . A pharmaceutical composition of clause 1 being a and attention disorders , gastroenteritis , duodenitis , jejunitis , capsule comprising a population of minibeads which have a ileitis, peptic ulcer, Curling ' s ulcer, appendicitis , colitis , diameter of at most 10 mm and which comprise the water pseudomembraneous colitis , irritable bowel syndrome e . g . soluble polymer matrix material and a coating on the matrix irritable bowel syndrome diarrhea predominant ( IBS - D ) , material, the water- soluble polymer matrix material further irritable bowel syndrome constipation predominant ( IBS including the hydroxylase inhibitor , wherein the hydrophilic C ) or irritable bowel syndrome mixed ( IBS - M ) ; diverticu surfactant has an HLB value of at least 15 , and wherein the losis , diverticulitis , endometriosis , colorectal carcinoma, coating comprises a controlled -release polymer , optionally adenocarcinoma, and chronic heart failure . wherein the coating is a barrier membrane for extended 43 . A composition of any of clauses 40 to 42 which further release of the water- insoluble active ingredient and of the comprises, or is for use in combination therapy with , another hydroxylase inhibitor and / or is a coating which resists therapeutically active agent, for example a hydroxylase becoming degraded or becoming of increased permeability inhibitor . in the conditions of the GI tract above the colon but which 44 . A composition of any of clauses 40 to 43 which becomes degraded or of increased permeability in the con comprises gelatin as the water- soluble polymer . ditions of the colon101 . 45 . A composition of clause 44 in which gelatin is substan 33 . A pharmaceutical composition of clause 32 wherein the tially the only water - soluble polymer. minibeads further comprise in the polymer matrix part a non - ionic surfactant comprising a poly (oxyethylene ) group 46 . A composition of any of clauses 40 to 45 wherein the and the hydrophilic surfactant is an anionic surfactant. water - soluble matrix material is selected from a hydrocol 34 . A composition of any preceding clause wherein the loid , a non -hydrocolloid gum and chitosan and derivatives water - insoluble active ingredient is a macrolide immuno thereof. suppressant, and optionally is a cyclosporin , tacrolimus, an 47 . A composition of any of clauses 40 to 46 wherein the ascomycin or sirolimus , and particularly is ciclosporin . water- soluble polymer matrix material is in the form of a 35 . A pharmaceutical composition of clause 31 or clause 32 plurality ofminibeads having a diameter of not more than 10 which further includes the specific feature ( s ) recited in any mm , e . g . of not more than 5 mm . one or a permitted combination of clauses 2 to 4 or 9 to 29 . 48 . A composition of clause 47 wherein the minibeads are 36 . A composition of any preceding clause wherein the monolithic , optionally with layers thereon . hydroxylase inhibitor is selected from hydralazine, DMOG , 49 . A composition of clause 47 or clause 48 wherein the FG -4497 and FG4095 , and in particular is hydralazine . minibeads comprise a controlled -release polymer . 37 . A composition of any preceding clause which further 50 . A composition of clause 49 wherein the controlled comprises another active pharmaceutical ingredient. release polymer is an extended release polymer or an enteric 38 . A composition of any preceding clause which comprises polymer. a gelatin capsule containing a plurality of minibeads into 51. A composition of clause 47 or clause 48 in which the which the water- soluble polymer matrix material is formed . minibead has a coat which comprises the controlled release 39 . A product for use in manufacturing a composition of any polymer and optionally a polymer susceptible to degradation preceding clause , the product being a water - soluble polymer by bacterial enzymes . matrix material formulated as a minibead having a diameter 52 . A composition of clause 49 wherein the controlled of no more than 10 mm , e . g. of not more than 5 mm , in release polymer is ethylcellulose comprised in a coating on US 2018 / 0042989 A1 Feb . 15 , 2018 35 the minibead and optionally in association with an emulsi - 74 . A composition of clause 73 wherein the water - soluble fication agent , for example ammonium oleate . polymer matrix material further comprises the hydroxylase 53 . A composition of clause 52 wherein the ethylcellulose is inhibitor. also in association with a plasticizer , e . g . dibutyl sebacate or 75 . A pharmaceutical composition of clause 40 being a medium chain triglycerides . capsule comprising a population of minibeads which have a 54 . A composition of clause 52 or clause 53 wherein the diameter of at most 10 mm and which comprise the water coating further comprises polymer susceptible of degrada soluble polymer matrix material and a coating on the matrix tion by bacterial enzymes . material, the water - soluble polymer matrix material further 55 . A composition of clause 54 wherein the polymer sus including a hydrophilic surfactant which has an HLB value ceptible of degradation by bacterial enzymes is water of at least 15 , and wherein the coating comprises a con soluble , preferably pectin . trolled - release polymer, optionally wherein the coating is a 56 . A composition of any of clauses 40 to 55 wherein the barrier membrane for extended release of the water - in water - soluble polymer matrix material further contains a soluble active ingredient and of the hydroxylase inhibitor and /or is a coating which resists becoming degraded or hydrophilic surfactant having an HLB value of at least 10 . becoming of increased permeability in the conditions of the 57 . A composition of clause 56 wherein the hydrophilic GI tract above the colon but which becomes degraded or of surfactant has an HLB value of at least 15 , and optionally of increased permeability in the conditions of the colon . at least 18 . 76 . A pharmaceutical composition of clause 75 wherein the 58 . A composition of clause 56 or clause 57 wherein the minibeads further comprise in the polymer matrix part a hydrophilic surfactant is an anionic surfactant. non - ionic surfactant comprising a poly ( oxyethylene ) group 59. A composition of clause 58 wherein the anionic surfac and the hydrophilic surfactant is an anionic surfactant. tant has an HLB value of at least 30 . 77 . A composition of any of clauses 40 to 76 wherein the 60 . A composition of clause 59 , wherein the anionic surfac water - insoluble active ingredient is a macrolide immuno tant comprises or is an alkyl sulfate salt . suppressant and optionally is a cyclosporin , tacrolimus, an 61. A composition of clause 60 in which the alkyl sulfate salt ascomycin or sirolimus, and particularly is ciclosporin . is sodium dodecyl sulfate (SDS ) 78 . A pharmaceutical composition of any of clauses 75 to 77 62 . A composition of any of clauses 57 to 61 in which the which further includes the specific feature ( s ) recited in any water - soluble polymer matrix material further contains a one or a permitted combination of clauses 2 to 4 or 10 to 28 . non - ionic surfactant having an HLB value of at least 10 but 79 . A method of making the composition of any preceding less than that of the hydrophilic surfactant. clause which comprises mixing an oil phase with an aqueous 63 . A composition of clause 62 wherein the non - ionic phase comprising a water soluble polymer matrix material to surfactant comprises a poly ( oxyethylene ) group . form an emulsion and then causing the emulsion to solidify . 64 . A composition of clause 63 wherein the non - ionic 80 . A method of clause 79 in which the emulsion is formed surfactant comprises a glycerol polyethylene glycol ricino into droplets which are then exposed to a solidification leate . medium . 65 . A composition of any of clauses 40 to 64 which 81. An emulsion for use in manufacturing a composition of comprises a liquid lipid and a solventmiscible therewith , in clause 39, the emulsion comprising oil droplets dispersed in which solvent the water - insoluble active ingredient is an aqueous phase characterised in that the aqueous phase soluble . comprises a water- soluble polymer matrix material and in 66 . A composition of clause 65 when dependent on any of that the emulsion comprises a water -insoluble active ingre clauses 62 to 64 in which the ratio of liquid lipid to non -ionic dient selected from calcineurin inhibitors , mTor inhibitors, surfactant is in the range 1 - 4 : 1 by weight, optionally 1 . 2 - 3 . and macrolide immunosuppressants , a hydroxylase inhibitor 0 : 1 by weight. and a hydrophilic surfactant having an HLB value of at least 67 . A composition of clause 65 or clause 66 in which the 10. liquid lipid is a medium chain triglyceride composition , the 82. A pharmaceutical composition for oral administration , medium chain triglyceride( s ) being one or more triglycer obtainable by : ides of at least one fatty acid selected from Co- C12 fatty [0413 ] C ) mixing together at least the following mate acids. rials to form an emulsion : 68 . A composition of clause 67 wherein the liquid lipid is a [0414 ] i ) a water - insoluble active ingredient selected caprylic /capric triglyceride . from calcineurin inhibitors, mTor inhibitors , and 69 . A composition of any of clauses 65 to 68 in which the macrolide immunosuppressants ; oil phase ( oil droplets ) represents from 10 - 85 % by dry [0415 ] ii ) an aqueous phase comprising water and a weight of the composition . water- soluble polymer material; 70 . A composition of any of clauses 65 to 69 in which said [0416 ] iii ) a hydrophobic liquid ; solvent is miscible with both the liquid lipid and with water. [0417 ] iv ) a hydrophilic surfactant having an HLB 71 . A composition of any of clauses 69 to 70 in which the value of at least 10 ; solvent is 2 - ( 2 - ethoxy )ethanol . [0418 ] V ) optionally one or more excipients which are 72 . A composition of any of clauses 40 to 71 in which the miscible with or soluble in hydrophobic liquid to unit solid or minibead has a low water content. increase the solubility of the immunosuppressant in 73 . A composition of any of clauses 40 to 72 which is for the liquid , combination therapy with a hydroxylase inhibitor comprised wherein the water - insoluble active ingredient is soluble in in a pharmaceutical formulation adapted to release the the hydrophobic liquid when it is combined with any said inhibitor in the colon . one or more excipients ; and US 2018 / 0042989 A1 Feb . 15 , 2018 36
[ 0419 ] D ) formulating the emulsion into a pharmaceu Mixing of the Two Phases tical composition comprising a unit solid which com prises the emulsion in a dry state , wherein the compo [0425 ] Oil Phase and Gelatin Phase are mixed in a 1 : 9 sition is adapted to release the active ingredient at least weight ratio . The resulting solution is stirred at 60 -70° C . to into the colon and the active ingredient is for therapeu achieve a complete homogeneity, then the homogenous tic use in combination therapy with a hydroxylase solution is ejected through a single orifice to form droplets inhibitor. which fall into a cooling oil medium (Miglyol 810N ) at 83 . A composition of clause 82 wherein the mixing together 8 - 10° C . The nozzle size ( diameter ) may be from 0 . 5 to 3 . 5 comprises forming a clear solution of the active ingredient mm . in the hydrophobic liquid together with any said one or more excipients . Filtering and Drying of Beads 84 . A composition of clause 82 or clause 83 wherein the formulating comprises ejecting the emulsion through a [ 0426 ] After approximately 30 minutes, beads are recov single -orifice nozzle , e . g . having a diameter of from 0 . 5 - 5 ered from the cooling oil , centrifuged to eliminate excess oil mm , to form drops which are then caused or allowed to fall and then dried at room temperature . into a cooling oil and allowed to harden to form minibeads , after which the minibeads are recovered from the cooling oil 1 . 2 Hydralazine / Cyclosporin A Combined Beads and dried . 85 . A method for administering to a subject a water - in 1 / Preparation of Gelatin Phase ( Hydralazine Lower soluble active ingredient selected from calcineurin inhibi Concentration ) tors , mTor inhibitors and macrolide immunosuppressants ; [0427 ] Hydralazine (1 .51 % w /w ), SDS ( 1. 50 % ), D - Sorbi and a hydroxylase inhibitor , the method comprising : tol ( 2 .01 % ) and gelatin (17 .50 % ) are added to water (77 . [0420 ] orally administering a pharmaceutical composition 48 % ) under constant stirring . The temperature is gradually comprising a unit solid which comprises a water - soluble increased to 60 -70° C . to achieve complete dissolution of polymer matrix material in which matrix material are dis gelatin . persed the active ingredient, droplets of water - immiscible liquid in which the active ingredient is soluble and a hydrophilic surfactant having an HLB value of at least 10 , 2 /Preparation of Gelatin Phase (Hydralazine Higher the composition being adapted to release the active ingre Concentration ) dient in at least the colon ; and [0428 ] Hydralazine ( 2 . 99 % w / w ), SDS ( 1 .51 % ), D - Sorbi [0421 ] simultaneous, sequentially or separately adminis tol (1 . 99 % ) and gelatin ( 17 .46 % ) are added to water (76 . tering a hydroxylase inhibitor to the subject . 05 % ) under constant stirring. The temperature is gradually 86 . A method of treating a disorder of, or at least suspected increased to 60 -70° C . to achieve complete dissolution of of being associated with , a leaky intestinal epithelial barrier, gelatin . the method comprising orally administering an effective amount of a pharmaceutical composition comprising a 1. 3 Cyclosporin A Beads water - soluble polymer matrix material in which are dis persed a water- insoluble active ingredient selected from [0429 ] The cyclosporin beads are prepared following the calcineurin inhibitors , m Tor inhibitors and macrolide immu general procedure set out in section 1 . 2 immediately above nosuppressants , and droplets of water - immiscible liquid in but omitting the hydralazine , and following the general which the active ingredient is soluble , wherein the compo procedure of Example 5 . sition is adapted to release the active ingredient in at least the colon . Preparation of Oil Phase . EXAMPLES 10430 ] Transcutol HP (40 .71 % w / w ) , Cremophor EL ( 22 . 22 % ) and Miglyol 810 N (11 . 13 % ) are stirred at room [ 0422 ] The following examples use a mouse model of temperature until a clear solution is obtained . Cyclosporin is colitis which is described next. then slowly added (25 . 92 % ) and the oil phase is stirred at room temperature until all the Cyclosporin is dissolved Example 1 — Manufacturing of Minibeads 1 . 1 Hydralazine Beads Mixing of the Two Phases [0431 ] Oil Phase and Gelatin Phase are mixed in a 1 : 7 Preparation of Gelatin Phase. weight ratio . The resulting solution is stirred at 60 -70° C . to [0423 ] Hydralazine ( 1 .50 % w /w ), SDS (1 . 50 % ) , D - Sorbi achieve a complete homogeneity , then the homogenous tol (2 .01 % ) and gelatin ( 17 .50 % ) are added to water (77 . solution is ejected through a single orifice to form droplets 49 % ) under constant stirring . The temperature is gradually which fall into a cooling oil medium (Miglyol 810N ) at increased to 60 - 70° C . to achieve complete dissolution of 8 - 10° C . The nozzle size (diameter ) may be from 0 . 5 to 3 . 5 gelatin . mm . Preparation of Oil Phase . Filtering and Drying of Beads [0424 ] Transcutol HP (54 . 96 % w / w ) , Cremophor EL (30 . [ 0432 ] After approximately 30 minutes, beads are recov 01 % ) and Miglyol 810 N ( 15 . 04 % ) are stirred at room ered from the cooling oil solution , centrifuged to eliminate temperature until a clear solution is obtained . excess oil and then dried at room temperature . US 2018 /0042989 A1 Feb . 15 , 2018 37
Mouse Model of Colitis - continued Summary of Procedures Component [0433 ] The chosen animal model uses dextran sodium Cremophor EL 8 . 95 sulphate (DSS ) to induce inflammation of the colon , and Gelatin 47. 27 measures the effectiveness of test formulations on control SDS 3 . 86 D - Sorbitol 5 . 44 ling clinical manifestations of disease . Inflammation of the Surelease (solid content) 3 .50 colon is induced using 2 . 5 % DSS in drinking water. The Pectin 0 .07 induction of colitis is quantified by determining the disease activity index (DAI ) . See Clinicopathologic study of dextran sulphate sodium experimentalmurine colitis , Lab Invest, 69 , [ 0443] Cyclosporin/ Hydralazine formulation (Formula 238 - 49 . Cooper, H . S . , Murthy, S . N . , Shah , R . S . and tion A & C Sedergran , D . J. ( 1993 ) . [0444 ] Formulation A (high level of Hydralazine in the combination ) TABLE 1 Scoring of disease activity index . Component % Cyclosporine 9 . 30 Score Weight loss Stool consistency Blood in feces Hydralazine 7 .54 None Normal None Transcutol P 14 .61 1 - 5 % Miglyol 810N 4 . 01 5 - 10 % Loose Hemoccult + Cremophor EL 7 .97 ??? 10 - 20 % Gelatin 43 . 94 > 20 % Diarrhoea Gross bleeding D - Sorbitol 5 .01 SDS 3 . 79 Surelease ( solid content) 3 . 77 [0434 ] The disease activity index is calculated as the sum Pectin 0 .08 of scores of weight loss , stool consistency and blood in feces . [0445 ] Formulation C ( low level of Hydralazine in the [0435 ] Normal stool= formed pellets [043 ] loose stool= pasty and semiformed stool which combination ) do not stick to the anus [0437 ] diarrhoea = liquid stools that stick to the anus Component % Cyclosporine 9 .69 Procedures Carried Out Hydralazine 3 . 95 Transcutol P 15 .21 [0438 ] DSS Colitis: Miglyol 810N 4 . 16 [0439 ] DSS (Dextran sodium sulfate ; 2 . 5 % ) is present in Cremophor EL 8 . 31 the drinking water of the mice for the duration of the Gelatin 45 .84 experiment. This is a standard model for IBD , The Hydroxy D - Sorbitol 5 . 25 lase Inhibitor Dimethyloxalylglycine is Protective in a SDS 3 .93 Murine Model of Colitis Gastroenterology , Volume 134 , Surelease ( solid content) 3 . 59 Issue 1 , January 2008 , Pages 156 - 165 .el . Eoin P . Cummins, Pectin 0 .07 Fergal Seeballuck , Stephen J . Keely, Niamh E . Mangan , John J . Callanan , Padraic G . Fallon , Cormac T . Taylor . At the [0446 ] The following dosages were administered : end of the experiment, the mice are euthanized by standard Formulation B : - 0 .629 mg CyA per 2 bead dose cervical dislocation . To monitor disease progression , disease Formulation A : - 0 .651 mg CyA and 0 . 528 mg HyA per 2 activity index (DAI ) is determined as discussed above . bead dose 10440 ] Administration of Mini -Beads : Formulation C : ~ 0 .678 mg CyA and 0 . 277 mg HyA per 2 [0441 ] The Mini- beads are administered using a stainless bead dose . steel oral gavage needle Two mini - beads are lodged into the 10447 ]. In the “ low HyA level” minibeads of formulation free end of the gavage needle and are administered orally to C , the weight ratio of HyA :CyA was about 0 . 4 : 1 . In the the mice along with 0 . 1 ml saline using standard 1 ml “ high HyA level ” minibeads of formulation A , the weight syringe ratio of HyA :CyA was about 0 . 8 : 1 . [0448 ] In the Dextran Sulphate Sodium (DSS ) model of Example 2 — Animal Study 1 colitis, mice are exposed to DSS in their drinking water which induces a loss of intestinal barrier function with Bead Formulations subsequent mucosal inflammation . Weight loss and disease activity index are used as a measure of disease progression . [ 0442] Cyclosporin formulation (Formulation B ) [0449 ] The results are shown in FIGS . 1 and 2 . 0450 ] The weight loss and DAI suggest that: Component % [0451 ] Formulation A (high level of Hydralazine ) Cyclosporine 10 . 48 shows the highest degree of protection in DSS induced Mygliol 4 . 47 colitis . Transcutol 15 . 96 [0452 ] Formulation B is protective at the early stage until day 4 and then shows a downward trend . US 2018 / 0042989 A1 Feb . 15 , 2018 38
[0453 ] Bead C is not protective at the early stage of -continued induction of colitis . Component % w / w Histology D - Sorbitol 4 . 94 [0454 ] Colon sections of euthanized mice are stained and SDS 3 .69 examined microscopically. Slides of the sections are shown Surelease (Solid Content ) 9 . 22 in FIG . 3 . The histology slides also show that Formulation Pectin 0 . 19 A has the highest degree of protection in DSS induced colitis . [0459 ] Formulation E ( 8 . 7 % wt. g Hydralazine beads ) Example 3 — Animal Study 2 Component % w / w Bead Formulations Hydralazine 4 . 11 Transcutol P 16 .65 [0455 ] Formulation A ( 3. 8 % wt.g Hydralazine beads) Miglyol 810N 4 . 56 Cremophor EL 9 . 09 Gelatin 47. 97 Component % w / w D - Sorbitol 5 . 50 SDS 4 . 12 Hydralazine 4 . 31 Surelease (Solid Content) 7 . 84 Transcutol P 17. 44 Pectin 0 . 16 Miglyol 810N 4 .77 Cremophor EL 9 .52 Gelatin 50 . 23 [0460 ) Formulation F (Uncoated HyA /CyA beads) D - Sorbitol 5 . 76 SDS 4 .31 Surelease (Solid Content) 3 .59 Pectin 0 . 07 Component % w / w Cyclosporine 10 .06 Hydralazine 4 . 10 [0456 ] Formulation B ( 3 .8 % wt. g hydralazine /cyclosporin Transcutol P 15 . 79 A beads) Miglyol 810M 4 .32 Cremophor EL 8 .62 Gelatin 47. 58 D - Sorbitol 5 .45 Component % w / w SDS 4 . 08 Cyclosporine 9 .69 Hydralazine 3 .95 Transcutol P 15 . 21 [0461 ] The following dosages were administered : Miglyol 810N 4 . 16 Formulation A : - 0 . 259 mg HyA per 2 bead dose Cremophor EL 8 . 31 Gelatin 45 . 84 Formulation B : - 0 .678 mg CyA and 0 . 277 mg HyA per 2 D - Sorbitol 5 .25 bead dose SDS 3 .93 Formulation C : - 0 . 268 mg HyA per 2 bead dose Surelease ( Solid Content) 3 .59 Formulation D : ~ 0 .638 mg CyA and 0 .260 mg HyA per 2 Pectin 0 .07 bead dose Formulation E : - 0 . 247 mg HyA per 2 bead dose [0457 ] Formulation C (Hydralazine uncoated beads) Formulation F : ~ 0 .704 mg CyA and 0 . 287 mg HyA per 2 bead dose . [0462 ] The weight loss change and disease activity index Component % w / w results are shown in FIGS . 4 and 5 . Hydralazine 4 . 47 [0463 ] From the weight loss data and DAI bead B ( 3 . 8 % Transcutol P 18 . 10 wt. g HyA /CyA beads) has a better protective efficacy fol Miglyol 810N 4 . 95 Cremophor EL 9 . 88 lowed by D and E . Gelatin 52 . 14 [0464 ] Colon sections of euthanized mice are stained and D - Sorbitol 5 . 97 examined microscopically . Slides of the sections are shown SDS 4 . 47 in FIG . 6 . The histology slides show that Formulations B ( 3 . 8 % wt. g . HyD /CYA beads ) has the highest degree of [0458 ] Formulation D (10 % wt.g HyA / CyA beads) protection in DSS induced colitis followed by D and E & F . Comparative ExampleMonotherapy with Component % w / w Hydralazine Cyclosporine 9 . 11 [ 0465 ] Hydralazine is administered to mice orally or i. p in Hydralazine 3 . 71 the DSS colitis model and the results ( weight change and Transcutol P 14 .31 Miglyol 810N 3 .91 DAD are shown in FIGS. 7 and 8 . Cremophor EL 7 . 81 [0466 ] The administration of Hydralazine API (oral and Gelatin 43 . 11 i. p . administration ) did not lead to any improvement both in terms of DAI and weight loss in DSS model of colitis . US 2018 / 0042989 A1 Feb . 15 , 2018 39
Example 4 — Caco - 2 Monolayer Tests [0475 ] 2. Transcutol P , Cremophor EL and Miglyol 810 are added together and mixed . Ciclosporin is then [0467 ] Transepithelial electrical resistance ( TEER ) is added and mixed until it a clear solution is obtained . measured in 5 animal groups ( see legend to FIG . 9 ) and the (Oil phase ) results are shown in FIG . 9 . [ 0476 ] 3 . The aqueous and oil phases are mixed to form [0468 ] As DSS is a chemical irritant which causes the an emulsion as described in Example 1 . disruption of the Caco - 2 monolayer , the TEER values are [ 04771 4 . The emulsion is then fed ( via temperature lowest in the case of DSS 4 % . TEER values for the DSS 4 % controlled tubing ) from the emulsion vessel through a alone receded to less that 20 % indicating complete disinte vibrating single orifice nozzle . The nozzle size (diam gration of the mono - layer. eter) is 3 .4 mm . [ 0469 ] Hydralazine 30 uM and 60 uM shows significant [0478 ] 5 . Seamless beads are formed as the solution protection to barrier function to Caco - 2 cells grown on flows through the vibrating nozzle into a chamber of permeable inserts in DSS induced in - vitro model of colitis . constantly flowing medium chain triglyceride (Miglyol [0470 ] Also monitored are changes in HIF - la expression 810 ) as a cooling oil. The cooling oil is at temperature in Caco - 2 cells with dosage of different hydralazine con of 8 - 10° C . centrations . The results are shown in FIG . 10 . 10479 ) 6 . The minicapsules are then removed from the cooling oil and placed in a centrifuge to remove the Example 5 — Comparative Efficacy of Treatment excess oil. with Cyclosporin a (Ciclosporin ) Via Oral [ 0480 ] 7 . Following centrifugation , Drying is initiated Minicapsules or i. p . Administration of the Same with a set refrigeration temperature . When the beads Dose of Cyclosporin A in a Spontaneous Mouse are rotating freely within the drum , they are considered Model of Colitis to be dried . [0481 ] 8 . The ethyl acetate is used to wash the mini [0471 ] In this example , the terms “ Cyclosporin A ” and capsules (remove any excess oil etc . ). The beads are “ CyA ” both refer to the drug whose international non dried for a further 24 h . proprietary name is “ ciclosporin ” . Also in this example , the [ 0482 ] 9 . The dried beads are then sieved to remove term “ SmPill ” refers to a minicapsule formulation of the oversize and undersize beads. The beads after sieving disclosure . are sized 1 mm - 2 mm . [ 0483 ] 10 . Pectin is added to purified water in a stainless Manufacturing Materials steel vessel and mixed to obtain a solution . Once the [0472 ] The materials used to make coated bead formula pectin is fully dissolved , Surelease® is slowly added to tions are as follows: the vessel whilst mixing is continued . [0484 ] 11. The beads are coated until a weight gain of 3 % has been reached . % Ingredient Wt/ Wt Function Chemical Composition 1 . Experimental Methods Ciclosporin 10 .59 Active Cyclic polypeptide Ingredient 1 . 1 IL - 10 - / - Colitis Model . Transcutol P 16. 07 Solubiliser 2 - ( 2 - ethoxyethoxy Jethanol Cremophor EL 8 . 96 Non - ionic Macrogolglycerol ricinoleate [0485 ] Mice with a disruption in the Interleukin -10 gene surfactant ( polyethoxylated castor oil ) ( IL - 10 - 1 - ) spontaneously develop chronic colitis and col Miglyol 810N 4 .54 Oil Triglycerides of fractionated Cg and C1o plant fatty acids orectal adenocarcinomas , with the age of onset and the D - Sorbitol 5 . 47 Plasticiser A sugar alcohol severity of the disease in IL - 10 - 1 - mice being dependent on Sodium Dodecyl 3 . 84 Anionic Sodium Dodecyl Sulphate background mouse strain and the conditions that the animals Sulphate surfactant are housed in . The progressive chronic enterocolitis in Gelatin Type A 47 .61 Bead shell - Protein IL - 10 - / - mice can be partially ameliorated by treatment of Porcine forming agent young mice with exogenous IL - 10 , but once disease is COATING MATERIALS established in adult animals the disease is not reversible by cytokine treatment with 100 % of mice affected . IL - 10 - / Surelease ® 2 . 85 Rate Ethylcellulose Controlling mice are routinely used as a model to test the efficacy of new Release drugs or treatment strategies for inflammatory bowel disease Polymer in humans for example , screening the efficacy of probiotics . Pectin 0 . 06 Pore Polysaccharide [0486 ] In this project , 5 - 6 week - old female IL - 10 - 1 - mice , former on a C57BL /6J strain background , were purchased from Jackson Laboratories (USA ) . Mice were kept in individually ventilated and filtered cages under positive pressure . Mice Manufacturing Method were fed an irradiated diet and housed on irradiated bedding . Food and water were supplied ad libitum . Sentinel mice [0473 ] The method generally follows the procedure of were screened to ensure SPF status. All animal experiments Example 1 and may be summarised as follows : were performed in compliance with Irish Department of [0474 ] 1 . Sodium dodecyl sulphate and D - sorbitol are Health and Children regulations and were approved by an mixed with purified water. Gelatin is then added to this ethical review board . solution and gentle heat applied ( to increase the tem [ 0487 ] The comparative efficacy of conventional delivery perature to 60 -70° C . ) until the gelatin has dissolved . of a daily dose of Cyclosporin A , i. p . or orally , versus ( Aqueous phase ) delivery of the same dose via incorporation into a minicap US 2018 / 0042989 A1 Feb . 15 , 2018 40
sule was tested in IL - 10 - 1 - mice on a C57BL /6J back - [0493 ] Colon extracts were analysed for TNF - a , IL - 1B ground . Drug treatments were administered to mice during and IL - 17, as representative proinflammatory cytokines , the chronic progression from mild to moderate colitis . Drugs using ELISA as described above . Myeloperoxidase (MPO ) were delivered daily ( a CyA daily dose of 0 .75 mg ) , see enzymatic activity in the colons was analysed as a marker of Table 1 , to 7 - 8 week -old IL - 10 - ) – mice , coincident with the inflammation . Cytokines and MPO are expressed as pg, or on - set of mild disease , with treatment continued for a further units per mg colon protein , respectively . 6 weeks when overt moderate colitis , i . e . no mortalities but clinical sign of colitis , develop . Mice were checked daily for 1 . 7 Histopathological Analysis of Colons. rectal prolapse and / or bleeding ( blood in faeces ) and mice 10494 ] When mice were euthanised , a ~ 1 cm piece of the were weighed weekly. proximal colon was removed and fixed in 10 % formalde hyde - saline . Sections were prepared and stained with hae 1. 2 Cyclosporin A (CyA ) Treatment Regime. matoxylin and eosin , as described ( Smith , P . et al . 2007. J [ 0488 ] All drugs were provided by Sigmoid Pharma Immunol 178 : 4557 -4566 ) . All histological scoring was per ( Table 1 ). All compounds and experimental groups were formed in a blinded fashion by two observers independently . randomly alphabetically labeled . Throughout experiments Colon sections from mice were graded using a histological all data recording and analyses were performed in a blind index ranging from 0 to 4 , based on a scoring system used manner. The codes on boxes / groups were not broken until in previous studies on IL - 10 - / - mice (McCarthy , J. et al. after the data was collected . 2003 . Gut 52 : 975 - 980 ) . This index was based on the degree of epithelial layer erosion , goblet cell depletion , and inflam 1. 3 Disease Activity Index (DAI ) . matory cell infiltrate (0 = normal; 1 =minimal evidence of [0489 ] To quantify the severity of colitis , a disease activity inflammatory infiltrate ; 2 = significant evidence of inflamma index (DAI ) was determined based on previous studies of tory infiltrate ( cryptitis , crypt abscesses) ; 3 = significant evi colitis in IL - 10 - ) - mice (Danese , S ., M . et al. , 2007 Gut dence of inflammatory infiltrate with goblet cell depletion ; 56 : 855 - 862 ) . DAI was calculated for individual mice 4 = significant evidence of inflammatory infiltrate with ero weekly . The DAI was based on a score of 1 for each of sion of the mucosa ) . The maximum possible score was 4 . ruffled fur, occult blood and soft stools . Animals with diarrhea or rectal prolapse that required humane killing of 1. 8 Statistical Analysis. the animal had an additional score of 2 added . A score was [0495 ] Power calculations on data from our previous stud given for each parameter , with the sum of the scores used as ies in IL - 10 - / - mice (3 ), determined a sample size of at least the DAI. The maximum potential score was 5 . 7 will detect significant difference (P < 0 .05 ) between groups in the 6 - week regime used . ANOVA and Tukey -Kramer 1 . 4 Serum Amyloid A Analysis . Multiple Comparisons Test determined statistical differ [0490 ] In Ulcerative colitis and Crohn ' s disease patients ences between multiple groups. 2 . the levels of Serum Amyloid A ( SAA ) are correlative with clinical disease and histology scoring of colon inflammation Results . (Niederau , C ., F. Backmerhoff, and B . Schumacher. 1997 . Testing Comparative Activity of Different Methods of Hepatogastroenterology 44 : 90 - 107) . In IL - 10 -/ - mice SAA Delivery of CyA in the IL - 10 - - Mouse Model of levels are also predictors of disease severity (Wei , X . et al. Spontaneous Colitis . 2008. Clin Immunol 129: 211 -218 ). Blood from mice was recovered at termination and serum isolated . Sera from mice [ 0496 ] Female IL - 10 - 1 - mice, all 5 - 6 weeks of age , were were tested by ELISA (Life Diagnostics Ltd ) for SAA . randomized into groups and housed for two weeks before drug treatment started . Groups of mice were treated as 1 .5 Spleen Cytokine Analysis . described in Table 1 . Drugs were administered and mice [ 0491] To address effects of differentmethods of admin monitored daily . Animals were weighed weekly . istration of CyA on systemic cell immunity , cytokine pro duction from spleen cells was examined after ex vivo TABLE 1 restimulation . Spleenswere removed from 4 mice per group , Experimental compounds administered . Cages of mice were labeled 1 - 4 by code number , at termination . Single cell suspensions by random coding, with the researcher blinded to the code throughout were cultured in vitro in media - alone or nonspecifically the experiment. Mice were treated as indicated daily . stimulated with 2 . 5 ng /ml Phorbal 12 -myristate 13 - acetate SmPill TM with CyA ORAL (PMA , Sigma, UK ) and 250 ng/ ml ionomycin (Sigma , UK ). ( 0 . 25 mg CyA /bead ) 3 beads daily in 0 . 2 mls of PBS Tumour necrosis factor ( TNF ) - a , Interleukin ( IL ) - 1ß and 0 .75 mg CyA /daily SmPill TM without ORAL IL - 17 levels in supernatants were detected by ELISA using CYA ( Vehicle ) 3 beads daily in 0 . 2 mlof PBS commercial kits ( R & D Systems) . CyA INTRAPERITONEAL Sandimmun ® 0 .2 ml per day 1. 6 Colon Cytokines and Myeloperoxidase Detection . (50 mg/ ml solution ) 0 .75 mg CyA/ daily ??? ORAL [0492 ] In order to assess inflammatory immune responses Neoral ® 0 . 1 mls per day at the site of disease , colons were removed from mice and ( 100 mg/ ml oral solution ) 0. 75 mg CyA /daily after removal of tissue for histology , immediately snap Untreated frozen and stored at - 80° C . Colons were homogenized and processed as described (Smith , P . et al . 2007. J Immunol [0497 ] There was death of one mouse, in SmPII (CYA ) 178: 4557 - 4566 ) . group on day 21 of the experiment ( This mouse was eutha US 2018 / 0042989 A1 Feb . 15 , 2018 ?) nized following tracheal damage as a result of drug admin (CyA ) groups of mice had significantly lower (P < 0. 01 ) istration via gavage ) . There were no other mortalities during histology scores relative to CyA (Oral ) , but not other groups the course of the experiment. No adverse effects were seen (FIG . 19 ) in any animals . 10505 ] To address systemic effects of drug treatment in [0498 ] In all groups there was a progressive increase in IL - 10 - /- mice , the spleens were removed at termination weight gain of mice until the 4th - 5th week of the treatment from 4 mice per group , selected 1 - 4 in order from the regime; thereafter all groups ceased to gain weight, or had numeric coding system , and spleen cells were cultured in weight loss (FIG . 12 ) . At termination , week 6 , a trend of vitro . The production of the three cytokines that were relatively less weight loss in SmPill (CyA )- treated mice measured in the colon ( IL - 1B , IL - 17 and TNF - a ) were (FIG . 12 ) was evident, with these mice being significantly assessed in spleen cells that were unstimulated ( cultured in heavier ( P < 0 .01 - 0 . 001) than all other groups (FIG . 13 ) . media ) or non -specifically activated by PMA / I treatment. [0499 ] No animals developed overt morbidity , rectal pro Following in vitro stimulation spleen cells from CyA ( i .p . ) lapse or frank diarrhea during the 6 -week treatment regime. treated mice has significantly reduced production of TNF - a However, from day 37 mice in the untreated and SmPill and IL - 17 relative to cells from untreated and SmPill (Ve ( Vehicle ) groups had evidence of blood in their faeces. hicle ) - treated mice (FIG . 20 ) . There were non - significant Based on DAI scoring on day 42 when mice were culled , the reductions in IL - 17 and TNF - a release by cells from other SmPill (CYA ) had the lowest DAI scores (FIG . 14 ), relative CyA - treated groups . The trend for reduced cytokine release to other groups, with these animals predominately having from CyA ( i . p . ) - treated mice was reflected in IL - 1B produc soft stools or occult faecal blood . SmPill (CYA )- treated mice tion , but there were no statistical differences between groups had significantly ( P < 0 .05 ) lower scores relative to untreated (FIG . 20 ) . or SmPill ( Vehicle ) groups, with non -significantly lower DAI values then CyA ( 1. p . ) or CyA (Oral ) groups (FIG . 14 ) . Example 6 — Mouse Model of Fibrosis [0500 ] Levels of SAA were analysed in serum isolated at termination on day 42 . The untreated group of mice had the [0506 ] The chosen animal model uses dextran sodium highest SAA levels , with the lowest in SmPill (CyA ) - treated sulphate (DSS ) to induce inflammation of the colon , fol mice and then CyA (i . p . ) - treated mice (FIG . 15 ). The lower lowed by a recovery phase to allow the development of relative levels in CyA ( i. p ) and SmPill (CyA ) groups were fibrosis . The mice are treated with various formulations significant (P < 0 .01 and P < 0 .001 , respectively ) relative to during the recovery phase to evaluate the effectiveness of untreated mice, but not other groups (FIG . 15 ) . test formulations on controlling clinical manifestations of [ 0501] As IL - 10 - / - mice develop colorectal adenocarci disease . Inflammation of the colon is induced using 2 . 5 % nomas the numbers of these were recorded at autopsy . While DSS in drinking water. there were differences between groups in the mean number [0507 ] Measurement of induction of inflammation is by of adenocarcinomas detected in the colons, with CyA (i . p . ) determining the disease activity index (DAI ) . See Clinico and SmPill (CYA ) groups having relatively less, the differ pathologic study of dextran sulphate sodium experimental ence was not statistically significant ( FIG . 16 ) . It was also murine colitis , Lab Invest, 69 , 238 -49 . Cooper, H . S . , noted that the adenocarcinomas seen in both these groups Murthy , S . N . , Shah , R . S . and Sedergran , D . J . ( 1993 ) . were smaller relative to those observed in the other three groups of mice ; however, this was not formally quantified . TABLE 1 (0502 ] At autopsy the entire colon , excluding tissue Scoring of disease activity index. removed for histology , was removed and processed to quan tify MPO enzymatic activity , as a marker of inflammation . Score Weight loss Stool consistency Blood in feces Additionally , the levels of the pro - inflammatory cytokines, None Normal None IL - 13 , IL - 17 and TNF - a were also quantified . The colons of 1 - 5 % the untreated and SmPill (Vehicle ) mice had the highest 5 - 10 % Loose Hemoccult + MPO enzymatic activity, with non - significantly lower enzy 10 - 20 % matic activity in CyA (Oral ) -treated mice ( FIG . 17 ). There Ohnmt > 20 % Diarrhoea Gross bleeding was statistically lower MPO activity in both CyA ( i. p . ) and SmPill (CyA ) groups relative to untreated mice ( P < 0 .05 ) , [0508 ] The disease activity index is calculated as the sum but not with the other groups. of scores of weight loss , stool consistency and blood in (0503 ] Consistent with reduced MPO levels in the colons faeces. of mice treated with CyA (i . p . ) or SmPill (CYA ), the levels [ 0509 ] Normal stool= formed pellets of all three inflammatory cytokines in the colons of these mice were also reduced relative to other groups of animals [ 0510 ] loose stool= pasty and semiformed stool which ( FIG . 18 ) . Compared to the untreated animals , SmPill do not stick to the anus (CYA ) - treated mice had a significant reduction in TNF - a [ 0511] diarrhoea = liquid stools that stick to the anus. and IL - 1B ( P < 0 .05 ) , whereas treatment with CYA ( i . p . ) [ 0512 ] Colon weight: The colon is emptied of the fecal reduced the colon levels of IL - 1ß but not TNF - a ( FIG . 18A , matter and weight of each colon is recorded . B ). There was no statistical significant difference in IL - 17 [0513 ] Colon length : The length of each colon is recorded . levels detected in the colons between groups ( FIG . 18C ) . [0514 ] Colon Histology : Approximately 10 mm of mid [0504 ] The proximal colon was removed at termination colon is fixed in 10 % buffered formaline and paraffin embed and Haematoxylin and Eosin stained sections prepared . ded . 4 um sections were stained with Hematoxyline and Scoring of colons demonstrated comparable degrees of mild Eosin stain to determine the degree of inflammation and inflammation in all groups, with CyA (Oral ) having the Masson trichrome stain for evaluation of fibrosis . The greatest relative score (FIG . 19 ) . CyA ( i . p . ) and SmPill stained sections were examined under light microscopy. US 2018 / 0042989 A1 Feb . 15 , 2018
Procedures Carried Out comparing different formulated drug concentrations with 105151 DSS Induced Fibrosis : equivalent concentrations of unformulated drug ; activation [0516 ] DSS (Dextran sodium sulfate ; 2 . 5 % ) is present in of NFKB was measured through NRE - Luciferase in HeLa the drinking water of the mice for 5 days duration , on day cells after drug treatment. 5 the DSS was stopped in drinking water and the treatment [ 0531 ] 6 different concentrations for Hydralazine HCl and groups were treated with various test formulations for 14 6 different concentrations for Cyclosporin A were used . The days. A normal recovery group did not receive any drug . following formulations were made according to the proce [ 0517 ] This is a standard model for Fibrosis: Kenji Suzuki dure of Example 1 and dissolved to provide solutions et al Pathology international 2011 ; 61: 228 - 238 . At the end containing the 6 different concentrations of active : of the experiment, the mice are euthanized by standard [0532 ] HyA Formulation cervical dislocation . To monitor disease progression , disease activity index (DAI ) is determined as discussed above. Component (0518 ] Administration of Minibeads : % w / w [0519 ] The minibeads are administered using a stainless Hydralazine 4 .47 Transcutol P 18. 10 steel oral gavage needle . Two minibeads are lodged into the Miglyol 810N 4 . 95 free end of the gavage needle and are administered orally to Cremophor EL 9 . 88 the mice along with 0 . 1 ml saline using standard 1 ml Gelatin 52. 14 syringe . D - Sorbitol 5 . 98 10520 ] The following dosages were administered in beads SDS 4 .47 similar to those of Example 2 : [ 0521 ] CyA formulation : ~ 0 .629 mg CyA per 2 bead dose [ 0533] CyA Formulation [ 0522 ] HyA formulation : - 0 .708 mg HyA per 2 bead dose [0523 ] CyA /HyA formulation : ~ 0 .710 mg CyA and - 0 . 780 mg HyA per 2 bead dose . Component % w / w [0524 ] The results are shown in FIGS. 21 -23 . The histol Cyclosporine 10 . 87 ogy of the colonic tissue shown in FIG . 32 suggests that the Miglyol 810N 4 .64 Hya + Cya formulation containing a high level of hydralazine Transcutol P 16 . 55 is more protective in DSS - induced colitis than the formu Cremophor EL 9 . 28 lation containing a low level of hydralazine . Gelatin 49. 02 SDS 4 .00 Example 7 — Mouse Model of Colitis with High D - Sorbitol 5 .64 and Low Ratio HyA : Cya Combinations [0525 ] The same animal model is used and the same [ 0534 ] Combination procedure followed as in Example 1 . [0526 ] The following combination dosages of hydralazine Component % w / w and cyclosporin A were administered in combination beads Cyclosporine 10 .06 similar to those of Example 2 : Hydralazine 4 . 10 [ 0527 ] Low level of HyA : ~ 0 .730 mg CyA and ~ 0 . 550 mg Transcutol P 15 . 79 HyA per 2 bead dose Miglyol 810N 4 .32 Cremophor EL 8 .63 [ 0528 ] High level of HyA : ~ 0 .710 mg CyA and ~ 0 .780 mg Gelatin 47 .58 HyA per 2 bead dose . D - Sorbitol 5 .45 [0529 ] In the “ low level ” minibeads, the weight ratio of SDS 4 . 08 HyA :CyA was about 0 .75 : 1 . In the “ high level” minibeads , the weight ratio of HyA :CHA was about 1 . 1 : 1 . The results [0535 ] The solutions of formulated Hydralazine HCl and are shown in FIGS. 24 - 26 . The histology of the colonic Cyclosporin A ( Sigmoid Pharma ) were diluted at different tissues in FIGS . 33 - 36 suggests that the Hya + Cya formula concentrations with complete DMEM medium ( Dulbecco ). tion containing a high level of hydralazine is more protective Stock solutions of Hydralazine HCl and Cyclosporin A were in DSS - induced colitis than the formulation containing a low prepared by dissolving drug powder . Hydralazine HCl was level of hydralazine . FIG . 33 comprises slides showing the dissolved in complete DMEM medium (Dulbecco ), 1600 histology of healthy control specimens. FIG . 34 comprises ug/ mL . Cyclosporin A was dissolved in ethanol, 100 slides showing the histology of DSS control specimens . FIG . mg/mL . The solutions were then filtered using a 0 . 2 um 35 comprises slides showing the histology of specimens of microfilter and diluted with DMEM medium (Dulbecco ) to colonic tissue from mice treated with “ low level” hydrala equivalent concentrations of the formulated drug solutions . zine/ cyclosporin A combination minibeads . FIG . 36 com 10536 ) Hela cells were grown in 6 - wells plates in order to prises slides showing the histology of specimens of colonic have 60 - 70 % confluent wells . Each cell well was then tissue from mice treated with “ high level” hydralazine / transfected with 200 ng NRE - Luc DNA plasmid and 100 ng cyclosporin A combination minibeads . B -Galactin DNA plasmid using Lipofectamine transfection . After 24 hours , to each well (1 mL ) 1 mL of different Example 8 Hydralzine HCl and Cyclosporin A solutions were added . [ 0530 ] The aim of the experiment is to prove the stability The cells were incubated at 37° C . and 5 % CO , for 5 hours . of Hydralazine HC1 and Cyclosporin A formulated (mini - (0537 ) After incubation , HeLa cells were washed with bead ) form and determine a relation between dose of drugs cold PBS and incubated for 5 minutes at RT with 1x and NFKB activation . The experiment was performed by luciferase lysis buffer (Promega ). The lysate were then US 2018 / 0042989 A1 Feb . 15 , 2018 43 scraped from the well and resuspended into a 1 . 5 mL tube . minigel system (Biorad ) using a 7 . 5 % acrylamide stacking The lysed cells were then centrifugated for 5 minutes at gel. Electrophoresis was carried out at a voltage of 90 V for 12 ,000 rpm and RT. The supernatant was collected in a new 2 hours . A protein marker (BioLabs ) was used . tube . The lysate was then analyzed using an NRE Luciferase [0547 ] After protein separation , the gel was removed from assay in order to quantify the NFKB expression . The mea the chamber and placed in chilled transfer buffer. Whatman sured RLU values were then normalized using the B - galac paper and nitrocellulose membrane (Biorad ) were also tosidase assay. moistened in chilled transfer buffer . The blot - sandwich was [ 0538 ] The measurement of Luciferase was repeated 3 assembled with , first a Whatman paper, second the nitrocel times per each sample in order to have a statistically lulose membrane, then the gel and finally a second Whatman significant result and the RLU values were represented in a paper . The proteins were transferred onto the nitrocellulose column graphic with 95 % confidential interval. membrane using a tank transfer system ( Biorad ) placed on 10539 ). The results are shown in FIGS. 29 , 30 and 31 , ice at a constant voltage of 100 V for 90 minutes . After the where the legend " unform ” refers to unformulated active transfer, the membrane was stained with Ponceau S solution and the legend " form ” refers to formulated (minibead ) ( Sigma ) to confirm the successful transfer of the proteins. active . The concentrations which appear in FIG . 31 are The membrane was then washed in PBST for 5 minutes to expressed as the concentration of hydralazine . remove the Ponceau S solution . The membrane was blocked during 1 hour by incubation with 10 % milk (Not fat dry milk Example 9 ( Fluka ) in PBST) . Then the membrane was incubated with [0540 ] The aim of the experiment is to prove the stability the first antibody ( Purified Mouse Anti -Human HIF - 1a (BD of Hydralazine HCl and Cyclosporin A formulated (mini Transduction laboratories ) , concentration 1 /750 in 10 % bead ) form and determine a relation between dose of drugs milk ) over night at 4° C . The membrane was then washed 4 and HIF - la activation . The experiment was performed by times with PBST for 5 minutes followed by incubation with comparing different formulated drug concentrations with the secondary antibody ( Anti -Mouse IgG HRP Conjugate equivalent concentrations of unformulated drug ; activation (Promega ), concentration 1 /2000 in 10 % milk ) for one hour. of HIF -la was detected through Western Blot in CaCo2 The membrane was again washed 6 times in PBST for 5 cells after drug treatment. minutes. 3 mL of ECL reagent pro membrane were prepared . [0541 ] The minibead formulations made according to the The membrane was carefully dried , covered with ECLplus procedure of Example 1 . reagent on a clean surface and incubated for 5 minutes at [0542 ] 6 different concentrations of Hydralazine HCl and room temperature. The membrane was dried as before, 6 different concentrations of Cyclosporin A were used . The placed into a film case and exposed to a X -ray film with minibead formulations used were the same as in Example 8 . exposition time of 6 seconds. After development the mem [0543 ] Solutions of formulated (SmPill ) Hydralazine HCI brane was washed with water and stored in PBST at 4° C . and Cyclosporin A ( Sigmoid Pharma ) were diluted at dif (0548 ) The exposed filmswere analyzed by comparing the ferent concentrations with complete DMEM medium (Dul bands with the protein marker. The bands of HIF - la were becco ). Stock solutions ofHydralazine HCl and Cyclosporin identified according to its molecular weight of 120 kDa. A were prepared by dissolving drug powder . Hydralazine 105491. The cells were treated with hydralazine at concen HCl was dissolved in complete DMEM medium (Dulbecco ) , trations in ug /mL of 0 , 10 , 20 , 100 , 200 and 300. The results 1600 ug/ mL . Cyclosporin A was dissolved in ethanol, 100 of the Hydralazine treated cells in FIG . 37 show an increase mg /mL . The solutions were then filtered using a 0 . 2 um of HIF - la expression proportional to the drug concentration microfilter and diluted with DMEM medium (Dulbecco ) to between 0 and 100 ug /mL and a stabilization of the expres equivalent concentrations of the formulated drug solutions . sion between 100 and 300 ug/ mL in both formulated and [ 0544 Caco2 cells were grown in 6 -wells plates in order unformulated groups . These results show an up - regulation to have 60 - 70 % confluent wells . To each well ( 1 mL ) 1 mL of different Hydralazine HC1 and Cyclosporin A solutions of HIF - la related to Hydralazine concentration and a sta were added . The cells were incubated at 37° C . and 5 % CO2 bility of the formulated drug . for 5 hours . [ 0550 ] The cells were treated with cyclosporin A at con [ 0545 ] After incubation , cells were washed with cold PBS centrations in ug /mL of 0 , 5 , 10 , 25 , 50 and 100 . The and incubated for 5 minutes on ice with " whole cell lysis expression of HIF - la on cells treated with Cyclosporin A is buffer” ( M . Tumbuwala ). The lysate were then scraped form only slightly detectable at this exposure time indicating a the well and resuspended into a 1 . 5 mL tube . The lysed cells low influence of this drug on regulation of HIF -la expres were then centrifugated for 5 minutes at 12 ,000 rpm and 4° sion , as can be seen in FIG . 38 . C . The supernatant was collected in a new tube and the f0551 ] The cells were treated with a combination of proteins concentration were determined using Biorad pro cyclosporin A and hydralazine at concentrations in ug/ mL of tein assay kit ( Sigma ) . 5 uL of lysate of each sample were 0 , 5 , 10 , 25 , 50 and 100 expressed as the concentration of added in each well of a 96 well plate , then 25 uL of solution hydralazine. In relation to cells treated with both Hydrala A and 200 uL of solution B were added to the samples. The zine HCl and Cyclosporin A , the results in FIG . 39 show a absorbance was read at 595 nm with reference filter at 450 marked up - regulation of HIF - 1a at 25 , 50 and 100 ug /ml , nm and the concentration of protein was measured using a which is not at all found with the CyA treatment. The greater standard curve . Protein concentration of lysate samples was influence of Hydralazine HCl on the HIF - la activation is then normalized at 871 ug /mL . clearly recognizable . To note is that the signals of formu [0546 ] 5x Laemmli buffer (Laemmli ) was added to nor lated drug is slightly stronger of that of unformulated drug . malized proteins and the protein samples were denatured by This could indicate a better stability of the combined drugs heating at 100° C . for 10 minutes . Protein samples were then in the formulated form , thanks to the delivery in different loaded ( 20 uL / slot) and separated on a vertical dual - slab phases. US 2018 /0042989 A1 Feb . 15 , 2018 44
Remarks fibrotic intestinal disorder , the method comprising adminis [ 0552] Hydralazine increases HIF - la expression in a dose tering a hydroxylase inhibitor to the patient. dependent manner indicating that hydralazine may act as an 2 . The method of claim 1 , wherein the maintenance hydroxylase inhibitor (Novel Mechanism of Action for therapy is for a patient who has suffered from or is suffering Hydralazine : Induction of Hypoxia - Inducible Factor- 1 from an inflammatory intestinal disorder. alpha, Vascular Endothelial Growth Factor , and Angiogen 3 . The method of claim 1 , wherein the maintenance esis by Inhibition of ProlylHydroxylases . Helen J . Knowles, therapy is for a patient who has suffered from or is suffering Ya -Min Tian , David R . Mole, Adrian L . Harris . Circ . Res . from a fibrotic intestinal disorder. published online Jun . 10 , 2004 ) . Referring to Example 2 this 4 . The method of claim 1 , wherein the disorder is selected mechanism may provide an explanation for hydralazine from ( i ) inflammatory bowel disease and ( ii ) graft - versus induced protection in DSS in - vitro and DSS induced murine host disease following hematopoietic stem cell transplanta model of colitis. The beta - actin band indicates that equal tion . amounts of protein were loaded in each lane . 5 . The method of claim 4 , wherein the disorder is colitis . 10553 ] The formulations of Example 2 with the higher 6 . The method of claim 1 , wherein the hydroxylase HyA /CyA combination not only provide overall protection inhibitor is at least one hydrolase inhibitor selected from (both DAI and Wt Loss ) but confer more protection at an DMOG , hydralazine , FG -4497 , FG4095 , AGN -2979 , TM earlier stage. This implies that improved barrier function 6008 , TM 6089 , a siRNA against hydroxylases, and an may assist in the maintenance of intestinal epithelial func antisense therapeutic against hydroxylases. tions and protect from the damage induced by DSS. In so 7 . The method of claim 6 , wherein the hydroxylase doing, the irritation is reduced and therefore the intestinal inhibitor is hydralazine . mucosa is less likely to suffer from inflammation . Nothing 8 . The method of claim 1 , wherein the hydroxylase completely protects cells from DSS damage but the HyA inhibitor is in solution phase in a pharmaceutical composi protection is an important phenomenon . Without being tion bound by theory , it is envisaged that, when the barrier 9 . The method of claim 8 , wherein the pharmaceutical function has been compromised , the presence of CyA will composition is a multiple minibead composition , the mini reduce inflammation . beads comprising a water soluble polymer matrix in which [ 0554 ] The observations of Example 2 are supplemented the hydroxylase inhibitor is in solid solution . by those of Example 9, showing HyA and HyA / CyA but not 10 . The method of claim 9 , wherein the minibeads further CyA alone up -regulate HIF - la expression . comprise a hydrophobic dispersed phase in the matrix , the [0555 ] The above is supported by the TEER observations. hydrophobic phase comprising a water immiscible liquid As noted , the DSS causes severe damage to the epithelial and being formed of material liquid at body temperature . barrier . Hy , provides protection from this damage . 11 . The method of claim 8 , wherein the pharmaceutical [ 0556 ] Importantly , HyA is much more effective when composition is adapted for at least a portion of the hydroxy delivered from the intestinal lumen by a minibead than when lase inhibitor to be released in the colon . administered orally or administered by i. p . injection . 12 . The method of claim 9 , wherein at least some of the [ 05571 Overall , the combination treatment both protects minibeads have a controlled release coating adapted for the the epithelial barrier and suppresses inflammation when coated minibeads to release hydralazine in the colon . administered locally . 13 . The method of claim 12 , wherein the coated mini 10558 ] However, monotherapy with cyclosporin is also beads are coated with a coating comprising a pH indepen shown to be effective in controlling manifestations of colitis dent polymer . and is indicated as potentially therapeutically effective 14 . The method of claim 1, wherein the disorder is ( alone or in combination therapy ) to treat disorders in which selected from celiac disease , HIV or another enteropathy, a “ leaky intestine ” is, or may be , implicated . pouchitis and cachexia . ( 0559] Example 5 shows that local delivery of a macrolide 15 . The method of claim 1 , wherein the disorder is a immunosuppressant, in this case ciclosporin , to the colon disorder of, or at least suspected of being associated with , a achieves differential modulation of colonic and systemic leaky intestinal epithelial barrier. cytokines . In other words , such local delivery preferentially 16 . The method of claim 15 , wherein the disorder is inhibits cytokine expression in the colon to achieve a local selected from celiac disease , a rheumatic disorder, rheuma anti - inflammatory effect without the same undesirable sys toid arthritis , temporomandibular joint syndrome , type I temic immunosuppression achieved by comparative formu diabetes , multiple sclerosis , atopic dermatitis , psoriasis, a lations Sandimmun® and Neoral which do not have a chronic pain syndrome, fibromyalgia , chronic fatigue syn controlled release function . The coated beads used in the drome, depressive disorders , affective disorders and atten examples achieve colonic targeting by inclusion in a poly tion disorders , gastroenteritis , duodenitis , jejunitis , ileitis , mer coat of a polymer which degrades in the presence of peptic ulcer, Curling ' s ulcer, appendicitis, colitis , bacterial enzymes present in the colon , specifically pectin ; a pseudomembraneous colitis , irritable bowel syndrome, pH - independent polymer, specifically ethylcellulose is diverticulosis , diverticulitis , endometriosis , colorectal carci included in the coating also . The invention therefore con noma , adenocarcinoma , and chronic heart failure . templates the use of dosage units having a polymer coating 17 . The method of claim 1 , wherein the method is a comprising a delayed release polymer and a polymer which method of reducing chemotherapy - induced or radiation degrades in the presence of bacterial enzymes present in the therapy - induced gastrointestinal insult and inflammatory colon . bowel disease , and combinations thereof. We claim : 18 . Themethod of claim 1 , wherein the patient is suffering 1 . A method ofmaintenance therapy of a patient who has from Crohn ' s disease , ulcerative colitis , diversion colitis , suffered from or is suffering from an inflammatory and/ or ischemic colitis , infectious colitis , chemical colitis , micro US 2018 / 0042989 A1 Feb . 15 , 2018 45 scopic colitis , atypical colitis , pseudomembraneous colitis , 31 . The method of claim 29 in which the water - soluble fulminant colitis , autistic enterocolitis, interdeminate colitis , polymer matrix material further contains a non - ionic sur Behcet' s disease , jejunoiletis , ileitis , ileocolitis , granuloma factant having an HLB value of at least 10 but less than that tous colitis , irritable bowel syndrome, celiac disease , stom of the hydrophilic surfactant. ach ulcers , diverticulitis , pouchitis , proctitis , mucositis , 32 . The method of claim 31 wherein the non - ionic sur radiation -associated enteritis , short bowel disease , or factant comprises a glycerol polyethylene glycol ricinoleate , chronic diarrhea. or another non - ionic surfactant comprising a poly (oxyeth 19 . Themethod of claim 1 , wherein the patient is suffering ylene) group . from collagenous colitis , lymphocytic colitis , and irritable 33 . The method of claim 10 in which the water - immis bowel syndrome with constipation , irritable bowel syn cible liquid comprises a liquid lipid and a solvent miscible drome with diarrhea and /or irritable bowel syndrome with therewith , in which solvent the water - insoluble active ingre pain symptoms. dient is soluble. 20 . A method of treating or preventing a disorder of a 34 . The method of claim 33 in which the water - soluble leaky intestinal epithelial barrier, wherein the disorder is not polymer matrix material further contains a non -ionic sur irritable bowel disease , inflammatory bowel disease , factant having an HLB value of at least 10 but less than that Crohn ' s disease, ulcerative colitis , GVHD , or GI- GVHD , of the hydrophilic surfactant and wherein the ratio of liquid themethod comprising administering to a patient a pharma lipid to non - ionic surfactant is in the range 1 - 4 : 1 by weight. ceutical composition for oral administration , comprising a 35 . The method of claim 33 in which the liquid lipid is a water -soluble polymer matrix material in which matrix medium chain triglyceride composition , the medium chain material are dispersed at least one water- insoluble active triglyceride ( s ) being one or more triglycerides of at least one ingredient selected from calcineurin inhibitor, m Tor inhibi fatty acid selected from C6 -C12 fatty acids . tor, macrolide immunosuppressant and macrolide antibiotic ; 36 . The method of claim 33 in which the solvent is and droplets of a water -immiscible liquid in which the 2 - ( 2 -ethoxy ) ethanol, or another solvent miscible with both water - insoluble active ingredient is soluble , wherein the the liquid lipid and with water . composition is adapted to release the water - insoluble active 37 . Themethod of claim 13 wherein the water- insoluble ingredient in at least the colon . active ingredient is comprised in a pharmaceutical formu 21 . A method according to claim 20 wherein the disorder lation adapted to release the active ingredient in at least the is selected from celiac disease , a rheumatic disorder, rheu colon . matoid arthritis , temporomandibular joint syndrome, type I 38 . The method of claim 20 , wherein the composition is diabetes, multiple sclerosis , atopic dermatitis , psoriasis, a a capsule comprising a population of minibeads which have chronic pain syndrome, fibromyalgia , chronic fatigue syn a diameter of at most 10 mm and which comprise the drome, depressive disorders , affective disorders and atten water- soluble polymer matrix material and a coating on the tion disorders , gastroenteritis , duodenitis , jejunitis , ileitis , matrix material, the water - soluble polymer matrix material peptic ulcer , Curling ' s ulcer , appendicitis , colitis , further including the hydroxylase inhibitor, wherein the pseudomembraneous colitis , diverticulosis , diverticulitis , hydrophilic surfactant has an HLB value of at least 15 , and endometriosis , colorectal carcinoma, adenocarcinoma, and wherein the coating comprises a controlled - release polymer , chronic heart failure . optionally wherein the coating is a barrier membrane for extended release of the water - insoluble active ingredient and 22 . The method of claim 9 , wherein the composition of the hydroxylase inhibitor and /or is a coating which resists comprises gelatin as the water - soluble polymer. becoming degraded or becoming of increased permeability 23. The method of claim 9 wherein the water - soluble in the conditions of the GI tract above the colon but which matrix material is selected from a hydrocolloid , a non becomes degraded or of increased permeability in the con hydrocolloid gum , chitosan and a derivative thereof. ditions of the colon . 24 . The method of claim 9 wherein the unit solid is a 39 . The method of claim 20 wherein the water- insoluble minibead having a diameter of not more than 10 mm , the active ingredient is a macrolide immunosuppressant. composition optionally comprising a plurality of said mini 40 . A multiple minibead unit dosage form , wherein the beads. minibeads comprise a water soluble polymer matrix com 25 . The method of claim 24 wherein the one or more prising hydralazine dispersed in the matrix . minibeads comprise a controlled -release polymer . 41. The unit dosage form of claim 40 , further comprising a dispersed phase composed of a material selected from a 26 . The method of claim 25 in which the minibead has a hydrophobic material , an amphiphilic material, and a com coat which comprises the controlled release polymer . bination thereof, the unit dosage form comprising hydrala 27. The method of claim 25 wherein the controlled zine in an amount of from 2 . 5 mg - 100 mg. release polymer is ethylcellulose comprised in a coating on 42. The dosage form of claim 40 wherein the hydralazine the minibead and optionally in association with an emulsi is in an amount of from 5 mg- 100 mg. fication agent. 43. The dosage form of claim 40, wherein the hydralazine 28 . The method of claim 8 wherein the composition is in solid solution in the matrix . further comprises a hydrophilic surfactant. 44 . The dosage form of claim 40 which is for once a day 29 . The method of claim 28 , wherein the hydrophilic administration . surfactant is an anionic surfactant. 45. A method of maintenance therapy by monotherapy 30 . Themethod of claim 29 , wherein the anionic surfac with the hydralazine , the method comprising administering tant comprises sodium dodecyl sulfate or another alkyl to a patient a multiple minibead unit dosage form according sulfate salt. to claim 38 . US 2018 /0042989 A1 Feb . 15 , 2018
46 . A method ofmaintenance therapy of a patient who has 59 . The method of claim 55 , wherein the method is suffered from or is suffering from an inflammatory intestinal maintenance therapy of a patient who has suffered from or disorder , themethod comprising administering to the patient is suffering from an inflammatory intestinal disorder , is the a multiple minibead unit dosage form wherein the minibeads prevention or reduction of chemotherapy - induced or radia comprise a water soluble polymer matrix in which a water tion therapy - induced gastrointestinal insult and inflamma soluble hydroxylase inhibitor is in solid solution . tory bowel disease , and combinations thereof, or the method 47 . The method of claim 46 , wherein the minibeads is the treatment of a patient suffering from Crohn ' s disease , further comprise a hydrophobic phase dispersed in the ulcerative colitis , diversion colitis , ischemic colitis , infec matrix , the hydrophobic phase being formed of material tious colitis , chemical colitis , microscopic colitis, atypical liquid at body temperature . colitis , pseudomembraneous colitis , fulminant colitis , autis 48 . A method of treating or delaying the progression of, a tic enterocolitis , interdeminate colitis , Behcet ' s disease , fibrotic intestinal disorder of a patient, the method compris - jejunoiletis, ileitis, ileocolitis , granulomatous colitis , irri ing administering a pharmaceutical composition comprising table bowel syndrome, celiac disease , stomach ulcers , diver a hydroxylase inhibitor to a patient in need thereof. ticulitis , pouchitis , proctitis , mucositis , radiation - associated 49 . The method of claim 48 , wherein the treatment is enteritis , short bowel disease, or chronic diarrhea . preventing or delaying the appearance of clinical symptoms of a state . 60 . The method of claim 55 , wherein the patient is 50 . The method of claim 48 , wherein the fibrotic disorder suffering from collagenous colitis , lymphocytic colitis , and is gastrointestinal fibrosis . irritable bowel syndrome with constipation , irritable bowel 51. The method of claim 48 , wherein the hydroxylase syndrome with diarrhea and /or irritable bowel syndrome inhibitor is in solution phase . with pain symptoms. 52 . The method of claim 48 , wherein the composition 61. The method of claim 55, wherein the weight ratio of comprises a polymeric matrix and the hydralazine is in hydroxylase inhibitor : immunosuppressant is from 0 . 8 : 1 to particulate form . 5 : 1 . 53 . The method of claim 48, wherein the hydroxylase 62. The method of claim 61, wherein the hydroxylase inhibitor is in solution phase in a pharmaceutical composi inhibitor is hydralazine and the immunosuppressant is tion ; wherein the pharmaceutical composition is a multiple cyclosporin . minibead composition , the minibeads comprising a water soluble polymer matrix in which the hydroxylase inhibitor is 63 . The method of claim 55 , wherein the immunosup in solid solution . pressant is selected from cyclosporin , tacrolimus, sirolimus , 54 . The method of claim 53 , wherein the minibeads pimecrolimus, an angiotensin II inhibitor, an ACE inhibitor, further comprise a hydrophobic dispersed phase in the a dicarboxylate - containing agent, a phosphate -containing matrix , the hydrophobic phase comprising a water immis agent, a casokinin , a lactokinin , a lactotripeptide , and / or the cible liquid and being formed of material liquid at body hydroxylase inhibitor is selected from DMOG , hydralazine , temperature . FG -4497 , FG4095 , AGN - 2979 , TM 6008 , TM 6089 , a 55 . A method of treating a warm - blooded animal to treat , siRNA against hydroxylases and an antisense therapeutic or delay the progression , of a fibrotic intestinal disorder, or against hydroxylases . for maintenance therapy of an animal which has suffered 64 . The method of claim 63 , wherein the hydroxylase from or is suffering from a fibrotic intestinal disorder, the inhibitor is in solution phase in a pharmaceutical composi method comprising administering to said animal simultane tion ; wherein the pharmaceutical composition is a multiple ously , sequentially or separately an immunosuppressant and minibead composition , the minibeads comprising a water a hydroxylase inhibitor. soluble polymer matrix in which the hydroxylase inhibitor is 56 . The method of claim 55 , wherein the treatment is preventing or delaying the appearance of clinical symptoms in solid solution . of a state. 65 . The method of claim 64 , wherein the minibeads 57 . The method of claim 55 , wherein the fibrotic disorder further comprise a hydrophobic dispersed phase in the is gastrointestinal fibrosis . matrix , the hydrophobic phase comprising a water immis 58 . The method of claim 55 , wherein the disorder is cible liquid and being formed of material liquid at body selected from celiac disease , HIV or another enteropathy, temperature . pouchitis , and cachexia . * * * * *