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USOO8354116B2

(12) United States Patent (10) Patent No.: US 8,354,116 B2 Carter et al. (45) Date of Patent: Jan. 15, 2013

(54) BIFUNCTIONAL SYNTHETIC MOLECULES Chinese Office Action, with English translation, dated Apr. 20, 2011 in corresponding foreign patent application No. CN 200880 103516. 1. (75) Inventors: Stephen G. Carter, Andover, MA (US); Wang, Xiu Q., et al., “Erythropoietin Depresses Nitric Oxide Kanu Patel, Londonderry, NH (US); Synthase Expression by Human Endothelial Cells'. Hypertension. Zhen Zhu, Tewksbury, MA (US); Lixin 1999, 33:894-899. Qiao, Tewksbury, MA (US) Krapf, Reto, et al., “Arterial Hypertension Induced by Erythropoietin and Erythropoiesis-Stimulating Agents (ESA). Clinical Journal of the American Society of Nephrology, Feb. 2009, vol. 4 No. 2, pp. (73) Assignee: BioChemics, Inc., Danvers, MA (US) 470-480 + refs. Canadian Communication dated Mar. 8, 2012 in corresponding (*) Notice: Subject to any disclaimer, the term of this Canadian patent application No. CA 2690357. patent is extended or adjusted under 35 Canadian Communication dated Jul. 25, 2011 in corresponding U.S.C. 154(b) by 1247 days. Canadian patent application No. CA 2690357. Chinese Communication, with English translation, dated Mar. 28. (21) Appl. No.: 11/820,172 2012 in corresponding Chinese Patent Application No. 200880 1035161, 11 pages. (22) Filed: Jun. 18, 2007 * cited by examiner (65) Prior Publication Data Primary Examiner — Robert A Wax US 2008/0312296A1 Dec. 18, 2008 Assistant Examiner — Melissa Mercier (51) Int. Cl. (74) Attorney, Agent, or Firm — Nields, Lemack & Frame, A6 IK9/00 (2006.01) LLC A6 IK 8/02 (2006.01) (52) U.S. Cl...... 424/400; 424/401 (57) ABSTRACT (58) Field of Classification Search ...... None The synthesis and use of bifunctional molecules to improve See application file for complete search history. the topical and transdermal delivery efficiency of various types of therapeutic agents or agents designed to promote the (56) References Cited transdermal delivery of those therapeutic agents either into the skin tissue or into the systemic circulation. Three major U.S. PATENT DOCUMENTS classes of molecules are covalently joined as bifunctional 4,555,398 A * 1 1/1985 Oda ...... 514,307 Substances; chemical vasodilators, passive dermal penetra 5,460,821 A 10, 1995 Masiz ...... 424/449 tion enhancers and therapeutic or diagnostic drugs. Chemical 5,645,854 A 7, 1997 Masiz ...... 424/449 vasodilators may be delivered into the skin to increasing the 5,853,751 A 12/1998 MasiZ ...... 424/449 blood flow in a tissue that has compromised circulation or

6,019,997 A * 2/2000 Scholz et al...... 424/449 6,372,712 B1 * 4/2002 Briesewitz et al...... 514, 20.9 they may be used as part of a delivery vehicle to promote the 6,635,274 B1 10/2003 Masiz ...... 424/449 delivery of the drug. Passive dermal penetration enhancers are those chemicals that promote the passive penetration of FOREIGN PATENT DOCUMENTS drugs and other chemicals through the stratum corneum and CA 2360590 A1 4/2002 epidermis of the skin tissue. Drugs and diagnostic agents are OTHER PUBLICATIONS the third group of chemicals that are candidates for the link age of molecules. Laietal. Structural Characteristics of Human Erythropoietin, Mar. 5, 1986. The Journal of Biological Chemistry, 3116-3121.* 4 Claims, No Drawings US 8,354,116 B2 1. 2 BFUNCTIONAL SYNTHETIC MOLECULES tional molecule with Superior dermal penetrating character istics combined with an active function or com BACKGROUND OF THE INVENTION bined with the drug. By covalently binding these agents together as they are formulated and maintaining this bond as The current invention describes the preparation of a novel they pass through the stratum corneum and epidermis, both series of chemically bifunctional function molecules with the functional groups are co-migrating in the same physical space design purpose of improving the pharmacologic characteris and also at the same time. The coordination of function and tics of either the vasodilator or the drug/diagnostic moiety. location in the skin tissue amplifies uptake of the drug or the There are published reports suggesting that in many medical delivery promoter, making the delivery vehicle more efficient conditions related to peripheral vascular disease and neuropa 10 and more substantial in effect. thy, including diabetic neuropathy, there is a compromised The combinations of individual molecules, as defined by microcirculatory environment around the nerves, which may classes according to function, include but are not limited to have an influence on the normal functions of the nerves, chemical vasodilators, passive dermal penetration enhancing leading to the abnormal sensations or lack of sensations noted agents and drugs or diagnostic agents. The identification of a with these medical conditions. Improving the metabolic State 15 reaction chemical functional group available to serve as a in the tissue around the nerve cells in the skin through an linkage point to bond with a different functional class of improved blood flow, increasing the oxygenation and nutrient molecule is critical to the selection of the specific molecules. delivery to the tissue as well as improving the removal of Examples of the chemicals available for selection for the cellular and tissue waste products, improves the health and chemical vasodilator class of molecules that are candidates to functioning of the nerve cells, which in turn creates an envi serve as one component of the bifunctional molecule, ronment for the reversal of a deteriorating medical condition include, by example only but are not limited to: , or slows the rate of deterioration. The use of bifunctional arginine, Sulphate, fumarate, benfu function molecules, including the use of vasodilators linked rodil hemisuccinate, benzyl nicotinate, hydro to a penetration enhancer molecule would improve the deliv chloride, hydrochloride, hydrochlo ery of these agents to the compromised tissue. 25 ride, citrate, , maleate, The preparation of the stable and covalently linked bifunc , di-isopropylammonium dichloroacetate, ethyl tional molecules from components such as a vasodilator (e.g., nicotinate, hepronicate, hexyl nicotinate, tartrate, nicotinic acid ortolaZoline) linked to a penetration enhancing nicotinate, hydrochloride, kallidinoge moiety (e.g., menthol or linoleic acid), allows for the more nase, methyl nicotinate, oxalate, nicametate cit efficient and coordinated delivery of the active agents (i.e., 30 rate, niceritrol, nicoboxil, nicofuranose, , drugs or vasodilators). The improved delivery of a functional nicotinyl alcohol tartrate, nitric oxide, nonivamide, oXpenti drug agent or vasodilator improves the treatment of a medical fylline, , papaveroline, , peroxynitrite, condition in a manner that can be more accurately measured pinacidil, pipratecol, propentofyltine, raubasine, , and predicted since the physicochemical characteristics and teasuprine, thymoxamine hydrochloride, tocopherol nicoti the temporal positioning of the molecule in the skin are better 35 nate, , nicotinate, diazoxide, hydralazine, understood and defined as opposed to those associated with minoxidil, and Sodium nitroprusside. Centrally acting agents an unlinked or un-bonded formulation containing the same include , quanaberZ, and methyl dopa. Alpha two Substances. adrenoceptor blocking agents include , phenoxy The efficiency and the focused application of use with the benzamine, , and . neuron present invention includes the use of a broad class of novel 40 blocking agents include bedmidine, debrisoquine, and molecules, containing at least two types of functional char . ACE inhibitors include benazepril, captopril, acteristics, including linking a vasodilator or a drug molecule cilaZapril, enalapril, fosinopril, lisinopril, perindopril, to a molecule designed to promote penetration through the quinapril, and ramipril. Ganglion-blocking agents include skin or to another type of vasodilator. The three classes of pentolinium and trimetaphan. Calcium channel blockers molecules: Vasodilators, penetration enhancers and drugs 45 include amlodipine, diltiazem, felodipine, isradipine, nicar may be linked as combinations of vasodilator to drug; vasodi dipine, nifedipine, nimodipine, and Verapamil. Prostaglan lator to penetration enhancer and penetration enhancer to dins including: prostacyclin, thrombuxane A2, leukotrienes, drug. PGA, PGA1, PGA2, PGE1, PGE2, PGD, PGG, and PGH. Angiotensin II analogs include Saralasin. Other Suitable DETAILED DESCRIPTION OF THE INVENTION 50 vasodilators include nitroglycerin, , thrazide, isosor bide dinitrate, pentaerythritol tetranitrate, digitalis, hydrala The invention describes the preparation of novel bifunc Zine, diaZOxide, and sodium nitroprusside. Typically the tional function molecules, created from chemical agents vasodilator linked to the penetration enhancer molecule, is known to participate in the promotion of transdermal drug present in the topical vehicle at concentration between delivery. In addition, the preparation of these bifunctional 55 0.0005% to 15% by weight, depending on the specific vasodi molecules may also be used for the enhanced delivery of one lator used and the pharmacologic properties of the chemical. of the components of the delivery vehicle, to serve as an active When the vasodilator is linked to a drug molecule, similar agent to itself. Such as a chemical vasodilator for the purpose concentrations are also used ranging from 0.0005% to 15% of improving local tissue blood flow in medical conditions by weight, with the specific concentrations on a mole basis known to exhibit compromised blood flow. 60 determined empirically by the bioavailability of the bifunc The basic design of this class of novel molecules combines, tional molecule. through a covalent or other formal chemical linkage, the Another functional component of a bifunctional molecule function chemical group of one of the classes described with is a dermal penetration enhancer. This class of molecules is the complimentary functional group or molecule on one of designed to assist in the transportation of the bifunctional the other classes of chemicals. 65 vasodilator molecule from the skin Surface, through the Stra The linkage of a penetration enhancer Substance with tum corneum and into the dermal layer of the skin. Suitable either a vasodilator or a drug molecule endows this bifunc penetration enhancers include by example only but are not US 8,354,116 B2 3 4 limited to: individual fatty acids, fatty acid esters, polyols, vasodilator, the penetration enhancer, or the drug molecule to amides, various anionic, cationic and nonionic Surfactants form the bifunctional molecule. One suitable functional Such as but not limited to Sodium laurate and sodium lauryl group is an acid halide. The following procedure is the pre sulfate, phospholipids, cholesterol and cholesterol deriva ferred synthetic procedure to convert either: vasodilators, tives, m-pyrrole, dimethyl acetamide, limonene, sphingolip penetration enhancers or drug molecules which contain an ids, ceramides, terpenes, alkanones, menthol, various organic acid functional group to acid halide intermediates, such as acids, Such as but not limited to salicylic acid, citric and acid chlorides, to allow the intermediate to serve as a reactant Succininc acid, prostaglandins, decyl methylsulfoxide, urea, for subsequent reactions in the development of bifunctional sulfoxide alcohols, plant extract oils. Suitable fatty acids molecules: include by example but are not limited to: linoleic acids, 10 linolenic acids, oleic acids, Stearic acids, and myristic acids. As an exemplary procedure to employ for the creation of Phospholipids include by example but are not limited to: bifunctional molecules between the different groups of phosphatidylcholine, phosphatidylethanolamine, and phos vasodilators, penetration enhancers, and drug molecules, the phatidylserine. Plant extract oils include peanut oil, hemp, first step in this process when the first molecule contains an borage, olive oil, Sunflower oil, Soybean oil, monoi oil and 15 acid functional group, is converted to an acid halide. Such as macadamia oil, with olive oil being preferred. Suitable alco an acid chloride. The following procedure outlines the gen hols for the plant extract oil/alcohol mix include ethyl alco eralized procedure for this purpose: hol, isopropyl alcohol, methyl alcohol. To a stirring mixture of a carboxylic acid-containing mol The third class of molecules that can be linked to either a vasodilator or a penetration enhancing molecule are thera ecule (1 mmol), pyridine (1.2 mmol) in 20 ml of dichlo peutic drugs and diagnostic agents. These drugs and agents romethane at 0°C. was added oxylyl chloride (1.2 mmol) and include all Small molecule, synthetic pharmaceuticals or 1 drop of anhydrous N,N-dimethylforman. The mixture was diagnostic agents as well as bio-therapeutic agents, which are then warmed to room temperature gradually, and the stirring typically protein or peptides, but may be members of other was continued for an additional 2 hrs. The resulting mixture classes of biological therapeutics, such as a lipid, carbohy 25 was evaporated to dryness, forming an acid halide. The acid drate or nucleic acid such as but not limited to insulin, anti halide residue was re-dissolved in 5 ml of anhydrous pyridine. sense oligonucleotides, lipopolysaccahrides, human growth 1 mmol of ROH or R'NH, was then introduced to the mixture hormone and erythropoietin. The common element for link and stirred at room temperature overnight. The reaction mix age of vasodilators or penetration enhancer molecules to each ture was diluted with 50 ml of ethyl acetate, washed with 20 of these small synthetic molecules or the biological therapeu 30 ml of 1.0MNaCO, aqueous solutions, and dried over anhy tics is that they all have functional chemical groups. Such as drous MgSO4. After concentration, the crude product was free and chemically reactive carboxylic acid or amine groups purified by flash column chromatograph on silica gel. The available for reacting with and forming a bifunctional mol result the synthesis of a bifunctional molecule between the ecule with various penetration enhancer and/or vasodilator first molecule containing a carboxylic acid group with the molecules. In addition to the functional groups on the thera 35 second molecule, containing a free hydroxyl or amino group. peutic or diagnostic agents being available chemically, the attachment of the vasodilator or penetration enhancer needs The starting materials used for the preparation of the to be either a chemically labile bond which may be cleaved bifunctional molecules with the vasodilators or drugs or pen upon entry into the body or a specific tissue or it needs to be etration enhancing chemicals listed by example or by infer physically positioned on the drug or diagnostic molecule Such 40 ence either naturally possesses the needed chemically reac that the function of the drug or diagnostic agent is not tive groups for the combination of the chemicals, or may be impaired. The satisfaction of this point needs to be evaluated modified to possess the needed chemically reactive groups. In in a bioassay to determine the drug or diagnostic agent activ Some instances, there may be a need or desire to prepare the ity before and after the delivery to the body. starting chemical in Such a way to create a more highly Examples of active ingredients that can be used in accor 45 reactive chemical or a specifically reactive chemical with the dance with the present invention include, but are not limited design to optimize the combination to form the bifunctional to: acetaminophen, acetohydoxamic acid, , or to direct the bifunctionalization in such a way to retain or acyclovir, albuterol, allopurinol, amiloride, amoxicillin, maintain the desired functionality of the bifunctionalized amphetamine, amplicillin, antisense polymers, . molecule once the combination has been completed. baclofen, beclomethasone, betamethasone, budesonide, 50 The following exemplify the preparation of bifunctional bumetanide, butorphanol, carbamazepine, carphenazine, molecules in accordance with certain embodiments. cefuroxime, cephradine, chloramphenicol, chlorothiazide, 1a) Bifunctional Molecule formation between a penetra chlorZoxazone, cinoxacin, cloraZepate, cloxacillin, cyclacil tion enhancer containing a hydroxyl group and a vasodi lin, dapsone, dicloxacillin, diethylstilbestrol, dopamine, lator containing a carboxylic acid group. Preparation of doxorubicin, erythropoietin, estradiol, fenoprofen, human 55 growth hormone, hydralazine, hydrochlorothiazide, ibupro analogs of menthol, as an example of a penetration fen, indomethacin, insulin, isoproterenol, levodopa, levothy enhancer molecule containing a hydroxyl group with a roXine, meclofenamate, melphalan, metformin methyl sali vasodilator chemical containing a carboxylic acid func cylate, metronidazole, minoxidil, morphine, . tional group Such as nicotinic acid: nalidixic acid, naproxen, nomifensine, norfloxacin, 60 oxaprozin, paramethasone, peptide fragments, , phenylpropanolamine, probenecid, quinethaZone, , O O Scopolamine, serotonin, , terfenadine, tocamide, triamterine, trimethoprim, and Valacyclovir. N OH (COCI) N C CH2C -- In accordance with certain embodiments, one of the 65 2 2 2 vasodilator, the penetration enhancer, or the drug molecule N Py, DMF N must have a functional group capable of reacting with a US 8,354,116 B2 5 6 -continued The result was the formation of a bifunctional molecule con N taining the vasodilator tolazoline and the penetration n enhancer oleic acid. Py 2) Examples of bifunctional molecules prepared combining He- 2 O 5 different chemical classes of passive penetration enhanc OH ing molecules reacted with different types of drugs con O taining different types of functional groups: 2a) Bifunctional Molecule formation between a therapeu 10 tic drug molecule containing a carboxylic acid group To a stirring solution of Nicotinic Acid (1 mmol) in 10 ml and a penetration enhancer molecule containing a free of dichloromethane at 0°C., was added oxylyl chloride hydroxyl group. Preparation of analogs of ibuprofen, as (1.1 mmol) and 5 ul of anhydrous N,N-dimethylfora an example of a drug containing reactive carboxylic acid mide (DMF). The resulting solution was warmed slowly groups with a penetration enhancer with a free alcohol to room temperature and the stirring was continued for 15 group Such as menthol: additional 2 hrs until no gas was evolved. After evapo ration of the solvent at reduced pressure, the residue was re-dissolved in 5 ml of anhydrous pyridine. 1 mmol of menthol was then added into the mixture and stirred at COOH (COCI) room temperature overnight. The reaction mixture was He diluted with 50 ml of ethyl acetate, washed with 20 ml of CH2Cl2 1.OMNaCO aqueous solutions, and dried over anhy Py, DMF drous MgSO. After concentration, the crude product was purified by flash column chromatograph on silica 25 COC Py gel. The result was the creation of a bifunctional mol -- He ecule between the vasodilator nicotinic acid and the penetration enhancer menthol. OH 1b) Bifunctional Molecule formation between a penetra tion enhancer containing a carboxylic acid group and a 30 vasodilator molecule containing an amine group. Prepa ration of analogs of oleic acid, as an example of a pen etration enhancer molecule containing a carboxylic acid group with a vasodilator with a reactive amine group Such as tolazoline: 35

(COCl) HC(CH2)6 1n o 1 (CH2)6COOH chool,CH2Cl2 Py, DMF 40 1n 1 (CH2)6COCl it To a stirring solution of Ibuprofen (1 mmol) in 10 ml of H3C(CH2)6 dichloromethane at 0°C., was added oxylyl chloride H N (1.1 mmol) and 5 ul of anhydrous N,N-dimethylfora Py He 45 mide (DMF). The resulting solution was warmed up N slowly to room temperature and the stirring was contin ued for additional 2 hrs until no gas was evolved. After evaporation of the Solvent at reduced pressure, the resi due was re-dissolved in 5 ml of anhydrous pyridine. 1 50 mmol of menthol was then added into the mixture and stirred at room temperature overnight. The reaction mix ture was diluted with 50 ml of ethyl acetate, washed with 20 ml of 1.0M NaCO aqueous solutions, and dried To a stirring solution of Oleic Acid (1 mmol) in 10 ml of dichloromethane at 0°C., was added oxylyl chloride (1.1 55 over anhydrous MgSO4. After concentration, the crude mmol) and 5ul of anhydrous N,N-dimethylforamide (DMF). product was purified by flash column chromatograph on The resulting Solution was warmed slowly to room tempera silica gel. The result was the creation of a bifunctional ture and the stirring was continued for additional 2 hrs until no molecule containing the active drug molecule ibuprofen gas was evolved. After evaporation of the solvent at reduced 60 and the penetration enhancer menthol. pressure, the residue was re-dissolved in 5 ml of anhydrous 2b) Bifunctional Molecule formation between active drug pyridine. 1 mmol of Tolazoline was then added into the mix molecules containing an amine group and penetration ture and stirred at room temperature overnight. The reaction enhancer molecules containing a carboxylic acid group. mixture was diluted with 50 ml of ethyl acetate, washed with Preparation of Analogs of Thiabendazole, as an 20 ml of 1.0M NaCO aqueous solutions, and dried over 65 Example of a Drug Containing Amine reactive groups anhydrous MgSO4. After concentration, the crude product with a penetration enhancer with a free carboxylic acid was purified by flash column chromatograph on silica gel. group Such as oleic acid: US 8,354,116 B2 7 8 ring was continued for additional 2 hrs until no gas (COCl) was evolved. The mixture was then evaporated to 1--"''' Py,CH2C, DMF b. Todryness. a stirring mixture of clonidine- 0 (1 mmol) in anhy (CH2)6COCl + 5 drous pyridine (5-10 mL), was added the freshly pre HccH1N=1 pared acid chloride (2 mmol). H c. The resulting mixture was then stirred at room tem N / S P perature overnight, and the pyridine was then evapo X-K - Y - rated under reduced pressure. e d. The residue was re-dissolved in Ethyl Acetate or N N o (CH2)6CH3 10 dichloromethane, and washed with Water. O 1 Yu- e. The organic layer was then dried over anhydrous Na2SO4. After concentration, the residue was puri N fied by re-crystallization in appropriate solvent. The N S residue contains the bifunctional molecule composed X-C 15 of clonidine and linoleic acid. A d Examples of acids linoleic acid, linolenic acid, oleic acid, etc. 3) Examples of bifunctional molecules prepared combining To a stirring solution of Oleic Acid (1 mmol) in 10 ml of different chemical classes of vasodilator molecules reacted dichloromethane at 0° C., was added oxylyl chloride 2 with different types of drugs containing different types of (1.1 mmol) and 5 ul of anhydrous N,N-dimethylfora functional groups: mide (DMF). The resulting solution was warmed up 3a) Preparation of analogs of thiabendazole as examples of drugs containing amine and Sulfur groups linked with slowly to room temperature and the stirring was contin chlorinated intermediates of carboxylic acid contain ued for additional 2 hrs until no gas was evolved. After 25 ingvasodilators such as nicotinic acid: evaporation of the Solvent at reduced pressure, the resi due was re-dissolved in 5 ml of anhydrous pyridine. 1 mmol of thiabendazole was then added into the mixture O and stirred at room temperature overnight. The reaction mixture was diluted with 50 ml of ethyl acetate, washed so N OH (COCI) N C with 20 ml of 1.0M NaCO aqueous solutions, and He- -- dried over anhydrous MgSO. After concentration, the Py,CH2C DMi 2 crude product was purified by flash column chromato- 4. H N graph on silica gel. The result was the creation of a N / S P bifunctional molecule containing thiabendazole and 35 X-O - Y - oleic acid with the respective properties of both on one N 2 molecule for the purpose of delivery characteristics. N 2c) Bifunctional Molecule formation between drug mol- Z \ ecules an amine group with a penetration enhancer in the form of an acid halide intermediate. Preparation of 40 O S. clonidine analogs, as an example of a drug containing double, or single amine groups in combination with the N S chlorinated carboxylic acid intermediate of a passive X-O dermal penetration enhancer Such as linoleic acid. 45 / 2

N C To a stirring solution of Nicotinic Acid (1 mmol) in 10 ml O of dichloromethane at 0°C., was added oxylyl chloride N ls (1.1 mmol) and 5 ul of anhydrous N,N-dimethylfora N + R C Pyridine 50 mide (DMF). The resulting solution was warmed up slowly to room temperature and the stirring was contin C ued for additional 2 hrs until no gas was evolved. After O R evaporation of the Solvent at reduced pressure, the resi N due was re-dissolved in 5 ml of anhydrous pyridine. 1 N C 55 mmol of thiabendazole was then added into the mixture and stirred at room temperature overnight. The reaction C X=N mixture was diluted with 50 ml of ethyl acetate, washed N with 20 ml of 1.0M NaCO aqueous solutions, and -N dried over anhydrous MgSO4. After concentration, the R o Cl 60 crude product was purified by flash column chromato graph on silica gel. The result was the formation of a bifunctional molecule composed of thiabendazole and a. To a stirring solution of Oleic Acid (1 mmol) in 10 ml nicotinic acid. of dichloromethane at 0°C., was added oxylyl chlo- 3b) Preparation of analogs as examples of ride (1.1 mmol) and 5 ul of anhydrous N,N-dimeth- 65 drugs containing carboxyl, amine and alcohol groups ylforamide (DMF). The resulting solution was with vasodilators containing carboxylic acids groups warmed up slowly to room temperature and the stir- Such as nicotinic acid: US 8,354,116 B2 9 10 (1.1 equiv) was added. The resulting solution was HO COOH stirred at room temperature overnight, and the pyri HC(OEt) --- dine was then evaporated under reduced pressure. The H2N CH F. residue was re-dissolved in ethyl acetate or dichlo HO romethane, and washed with water. The organic layer O COOEt was then dried over anhydrous NaSO. After concen R1COC EtO Her tration, the residue was purified by re-crystallization H2N CH Py in appropriate solvent. The product is amide 2. O HCI 4. Amide 2 was dissolved in anhydrous MeOH, and 2.0 10 1 Maqueous LiOH (2.0 equiv) was added. The solution O COOEt 1) LiOH, was stirred at room temperature for 2 hr, and neutral MeOH-HO ized with diluted HCl solution to pH-7. After evapo EtO -- HN CH 2) HCI ration, the residue was extracted using dichlo romethane-H2O. The organic layer is dried and O > 15 O R1 concentrated to give acid 3. 2 5. Acid 3 was converted to corresponding acid chloride O COOH using the same method in step 1. To the alcohol R2OH 1) (COCl) in pyridine, was added the corresponding acid chlo EtO He ride derived from acid 3. The resulting solution was HN CH stirred at room temperature overnight, and the pyri O 2n dine was then evaporated under reduced pressure. The O R1 residue was re-dissolved in ethyl acetate or dichlo romethane, and washed with water. The organic layer O COOR2 MeOH-H2O, 25 was then dried over anhydrous NaSO. After concen TSOH tration, the residue was purified by re-crystallization EtO HN CH3 rt, 16 hr in appropriate solvent. The product is ester 4. O > . Ester 4 was dissolved in MeOH HO (v/v 1/1), O R1 para-tosylsulfonic acid (0.1 equiv) was added. The 30 resulting solution was stirred at room temperature overnight, and evaporated to dryness. The resulting HO COOR catechol 5 was extracted using EtOAc—H2O, and R3COC re-crystallized in proper solvent, HN CH Py 7. R3COOH was first converted to the desired acid chlo HO 2n 35 ride using method in step 1. O R1 8. To a solution of catechol 5 in pyridine, was added the acid chloride R3COCl. After overnight stirring, the pyridine was evaporated under reduced pressure. The residue was re-dissolved in ethyl acetate or dichlo 40 romethane, and washed with water. The organic layer O COOR2 was then dried over anhydrous NaSO. After concen tration, the residue was purified by re-crystallization HN CH in appropriate solvent. The final product is bifunc tional molecule composed of methyldopa and two 45 different penetration enhancer components. n > 9. Examples of RCOOH, RCOOH-linoleic acid, lino lenic acid, oleic acid. 10 ROH-menthol, HO(CH), CH=CHCH-CH=CHCHCH-CH(CH), CH, 1. Method to convert an acid-containing active drug 50 HO(CH),CH=CH(CH),CH3 molecule Such as methyldopa to an acid chloride: To 3c) Ibuprofen reacted with the vasodilator tolazoline as an a stirring solution of methyldopa acid (1 mmol) in 10 example of a drug containing a free carboxylic acid ml of dichloromethane at 0°C., was added oxylyl group combined with an amine-containing vasodilator: chloride (1.1 mmol) and 5ul of anhydrous N,N-dim- 55 ethylforamide (DMF). The resulting solution was warmed up slowly to room temperature and the stir ring was continued for additional 2 hrs until no gas was evolved. The mixture was evaporated to dryness. COOH (COCI) 2. To the solution of methyldopa (1 equiv) in anhydrous 60 CH2Cl2 ethanol, was added ethyl orthoformate (3 equiv) and Py, DMF HC1-dioxane (commercially available from Aldrich, 1.5 equiv). The resulting mixture was refluxed for 4 hrs, and evaporated under reduced pressure to yield COC compound 1. 65 3. Compound 1 was dissolved in anhydrous pyridine, and freshly prepared acid chloride according step 1 US 8,354,116 B2 11 12 -continued 4) The resulting acid was converted to corresponding H acid chloride using same procedure in step 1, and N Py react with R2OH -in pyridine.- - - N ) - Y - 5) The resulting- solution- - - was stirred at room temperature overnight, and the pyridine was then evaporated under O reduced pressure. The residue was re-dissolved in ethyl acetate or dichloromethane, and washed with N water. The organic layer was then dried over anhy X 10 drous NaSO4. After concentration, the final product N is obtained after flash column chromatography. Examples of RCOOH-linoleic acid, linolenic Acid, oleic acid. ROH-menthol, HO(CH), To a stirring solution of Ibuprofen (1 mmol) in 10 ml of CH=CHCH-CH=CHCH-CH=CH(CH),CH, dichloromethane at 0°C., was added oxylyl chloride 15 HO(CH),CH=CH(CH),CH3 (1.1 mmol) and 5 Jul of anhydrous N,N-dimethylfora 3e) Preparation of drugs containing reactive amine groups mide (DMF). The resulting solution was warmed up slowly to room temperature and the stirring was contin Such as lisinopril and quinapril, with carboxylic acid ued for additional 2 hrs until no gas was evolved. After containing vasodilators, such as nicotinic acid, the evaporation of the solvent at reduced pressure, the resi 2O method used to prepare this novel class of analogs is due was re-dissolved in 5 ml of anhydrous pyridine. 1 identical to the method described in 3c. mmol of tolazoline was then added into the mixture and stirred at room temperature overnight. The reaction mix O ture was diluted with 50 ml of ethyl acetate, washed with 25 20 ml of 1.0M NaCO aqueous solutions, and dried ls over anhydrous MgSO4. After concentration, the crude HN R1 product was purified by flash column chromatograph on silica gel. 3d) Preparation of captopril-vasodilator bifunctional mol- so O ecules, as examples of combining drugs containing both RO carboxyl and thiol groups with a chlorinated intermedi- 2 ate of a vasodilator such as nicotinic acid. N R 35 -N O COOR O R1 O CO2H ~. s Lisinopril Analogs HS N RCOCI E -- CH aq. NaHCO3 40 ls --- CO2H R1 S E N 1. (COCl) O O2C, E 2. R2OH EtO R. ''. CH 45 O O N

R1 S N -N O R1 O CH3 50 Quinapril Analogs 1) To a stirring solution of oleic acid (1 mmol) in 10 ml of dichloromethane at 0°C., was added oxylyl chlo- 3f) Preparation of drugs containing both amine and alcohol ride (1.1 mmol) and 5 ul of anhydrous N,N-dimeth- ss groups such as phentolamine combined with vasodila ylforamide (DMF). The resulting solution was tors such as chlorinated intermediates of nicotinic acid. warmed up slowly to room temperature and the stir ring was continued for additional 2 hrs until no gas was evolved. The mixture was then evaporated to ".) dryness. 60 S. 2) With an ice bath, to the stirring suspension of capto- N pril (1 equiv) in the mixed solvent of THF-10% aq. RICOCI NaHCO (v/v=3/1, NaHCO2 equiv) was added the N OH aq. NaOH freshly prepared acid chloride in step 1 (1.2 equiv). 3) The mixture was stirred at 0°C. for 1 hr., and 10°C. for 65 1 hr, and then extracted with EtOAc. The organic layer H3C was separated, dried, and concentrated. US 8,354,116 B2 13 14 -continued Once the bifunctional functioning molecules have been O synthesized, they may be incorporated as part of a topical or transdermal drug delivery vehicle, which may take the form of a solution, Suspension, emulsion, lotion or cream or other R1 ) pharmaceutical formulation designed to be applied to the skin S. Surfaces. These formulations may be constructed as aqueous N or anhydrous based formulations depending on the chemical R2COCl N OH - - nature of the active and the Supporting components of the Py formulation to enable to these components to achieve maxi 10 mum functionality as a result of the chemical compatibility of those ingredients with chemical composition of the formula H3C tion. For example, a water-soluble or compatible bifunctional O molecule would be best suited to be prepared as part of a water-based formulation. In addition, these formulations may 15 contain a lipid enriched composition to assist the complex to ry penetrate through the stratum corneum as well as preferred S. N composition of a molecular occlusive barrier to promote the penetration and also to prevent the loss of water from the skin tissue. N "S R2 Example 1 of a bifunctional molecule incorporated into a O formulation composition as a lotion: HC 5% (w/w) Bifunctional molecule of a drug-vasodilator (i.e., ibuprofen-methyl nicotinate) 5% oleic acid 1) To a stirring solution of oleic acid (1 mmol) in 10 ml 25 5% linolenic acid of dichloromethane at 0°C., was added oxylyl chlo 3% propylene glycol ride (1.1 mmol) and 5 ul of anhydrous N,N-dimeth 2% pemulen ylforamide (DMF). The resulting solution was 2% lipomulse 165 warmed up slowly to room temperature and the stir 1% cetyl alcohol ring was continued for additional 2 hrs until no gas 30 1% allantoin was evolved. The mixture was then evaporated to 76% distilled water dryness. The bifunctional molecule and allantoin are dissolved into 2) With an ice bath, to the stirring suspension of phen the water for the formulation in one vessel. Once dissolved, tolamine (1 equiv) in the mixed solvent of THF-10% then the propylene glycol and pemulen are added slowly aq. NaOH (v/v=3/1, NaOH 2 equiv) was added the 35 while mixing vigorously. In a separate vessel, containing the freshly prepared acid chloride in step 1 (1.2 equiv). oleic acid and linolenic acid, the oil is heated to 55° C. then 3) The mixture was stirred and gradually warmed to the cetyl alcohol and lipomulse are added while mixing until room temperature for 2 hours. melted. The oil phase is allowed to cool to 30° C. then added 4) The mixture was neutralized to pH-7 and extracted slowly while stirring the water phase and blended until a 40 proper emulsion has been formed. with EtOAc. The organic layer was separated, dried, The bifunctional molecule may be delivered by directly and concentrated. applying the Substance to the skin and allowing the penetra 5) To a stirring solution of oleic acid (1 mmol) in 10 ml tion characteristics of the formulation to enhance the delivery of dichloromethane at 0°C., was added oxylyl chlo of the material into the skin tissue. Alternatively, the formu ride (1.1 mmol) and 5 ul of anhydrous N,N-dimeth 45 lations may also be administered with the assistance of an ylforamide (DMF). The resulting solution was external apparatus Such as but not limited to a sonophoretic or warmed up slowly to room temperature and the stir iontophoretic device, in a process designed to introduce the ring was continued for additional 2 hrs until no gas bifunctional complex and the drug or diagnostic molecule was evolved. The mixture was then evaporated to into the dermis. dryness. 50 Those skilled in the art will appreciate that the drug mate 6) The residue made in step 4 was dissolved in anhy rial used may serve a second function to the process, for drous pyridine, while freshly prepared acid chloride example the drug may be the vasodilator and as Such, the in step 5 was added slowly. After overnight stirring, creation of a vasodilator linked to a penetration enhancer the pyridine was evaporated under reduced pressure. molecule may be the objective for cases that are focused on a The residue was re-dissolved in ethyl acetate or 55 clinical endpoint of improving localized blood flow. dichloromethane, and washed with water. The organic layer was then dried over anhydrous NaSO. After What is claimed is: concentration, final product is obtained after flash 1. A method for the preparation of a bifunctional function column chromatography. ing molecule, comprising providing a first molecule having Examples of R1COOH, R2COOH=linoleic acid, oleic 60 one of vasodilatory or dermal penetration enhancing proper acid, linolenic acid. ties that has a first reactive group in the molecule selected The examples of the chemical entities, individually as well from the group consisting of an amine, an alcohol, a carboxy as the bifunctional combinations of those entities are listed lic acid functional group, and a hydroxyl group, and reacting and outlined above as examples only and are not limited to said first molecule with a second molecule having the other of only those chemicals but rather to serve as examples of 65 vasodilatory or dermal penetration enhancing properties that classes of molecules both in terms of functional group reac has a second reactive group in the molecule reactive with said tivity as well as in class of molecule. first reactive group, said second reactive group being selected US 8,354,116 B2 15 16 from the group consisting of an amine, an alcohol, a carboxy benazepril, captopril, cilaZapril, enalapril, fosinopril, lic acid functional group, and a hydroxyl group. lisinopril, perindopril, quinapril, ramipril, pentolinium, 2. A bifunctional molecule comprising trimetaphan, amlodipine, diltiazem, felodipine, israd a. a vasodilator selected from the group consisting of ipine, nicardipine, nifedipine, nimodipine, Verapamil, amrinone, arginine, bamethan Sulphate, bencyclane 5 prostacyclin, thrombuxane A2, leukotrienes, PGA, fumarate, benfurodil hemisuccinate, benzyl nicotinate, PGA1, PGA2, PGE1, PGE2, PGD, PGG, PGH, sarala buflomedil hydrochloride, buphenine hydrochloride, sin, nitroglycerin, labetalol, thrazide, isosorbide dini butalamine hydrochloride, cetiedil citrate, ciclonicate, trate, pentaerythritol tetranitrate, digitalis, hydralazine, cinepazide maleate, cyclandelate, di-isopropylammo diazoxide, and sodium nitroprusside; nium dichloroacetate, ethyl nicotinate, hepronicate, 10 b. an active drug molecule selected from the group consist hexyl nicotinate, ifenprodil tartrate, , ing of acetaminophen, acetohydoxamic acid, isoXSuprine hydrochloride, kallidinogenase, methyl acetophenazine, acyclovir, albuterol, allopurinol, nicotinate, naftidrofuryl oxalate, nicametate citrate, nic amiloride, amoxicillin, amphetamine, amplicillin, eritrol, nicoboxil, nicofuranose, nicotinyl alcohol, nic atenolol, baclofen, beclomethasone, betamethasone, otinyl alcohol tartrate, nitric oxide, nonivamide, Oxpen 15 budesonide, bumetanide, butorphanol, carbamazepine, tifylline, papaverine, papaveroline, pentifylline, carphenazine, cefuroxime, cephradine, chlorampheni peroxynitrite, pinacidil, pipratecol, propento?yltine, col, chlorothiazide, chlorZoxaZone, cinoxacin, cloraZe raubasine, Suloctidil, teasuprine, thymoxamine hydro pate, cloxacillin, cyclacillin, dapsone, dicloxacillin, chloride, tocopherol nicotinate, tolazoline, Xanthinol diethylstilbestrol, dopamine, doxorubicin, estradiol, nicotinate, diaZoxide, hydralazine, minoxidil, sodium fenoprofen, human growth hormone, hydralazine, nitroprusside, clonidine, quanaberZ, methyl dopa, hydrochlorothiazide, ibuprofen, indomethacin, insulin, indoramin, , phentolamine, pra isoproterenol, levodopa, levothyroxine, meclofenamate, Zosin, bedmidine, debrisoquine, guanethidine, melphalan, metformin methyl salicylate, metronidazole, benazepril, captopril, cilaZapril, enalapril, fosinopril, minoxidil, morphine, nadolol, nalidixic acid, naproxen, lisinopril, perindopril, quinapril, ramipril, pentolinium, 25 nomifensine, norfloxacin, oxaprozin, paramethasone, trimetaphan, amlodipine, diltiazem, felodipine, israd perphenazine, phenylpropanolamine, probenecid, ipine, nicardipine, nifedipine, nimodipine, Verapamil, quinethaZone, ritodrine, Scopolamine, serotonin, terb prostacyclin, thrombuxane A2, leukotrienes, PGA, utaline, terfenadine, tocainide, triamterine, trimethop PGA1, PGA2, PGE1, PGE2, PGD, PGG, PGH, sarala rim, and Valacyclovir, sin, nitroglycerin, labetalol, thrazide, isosorbide dini 30 wherein said vasodilator and said active drug molecule trate, pentaerythritol tetranitrate, digitalis, hydralazine, are covalently linked. diazoxide, and Sodium nitroprusside; 4. A bifunctional molecule comprising ... a dermal penetration enhancer selected from the group c. a. a dermal penetration enhancer selected from the group consisting of linoleic acids, linolenic acids, oleic acids, consisting of linoleic acids, linolenic acids, oleic acids, Stearic acids, and myristic acids; sodium laurate, sodium 35 Stearic acids, and myristic acids; sodium laurate, sodium lauryl Sulfate, phosphatidylcholine, phosphatidyletha lauryl Sulfate, phosphatidylcholine, phosphatidyletha nolamine, phosphatidylserine, cholesterol, m-pyrrole, nolamine, phosphatidylserine, cholesterol, m-pyrrole, dimethyl acetamide, limonene, menthol, Salicylic acid, dimethyl acetamide, limonene, menthol, Salicylic acid, citric acid, Succininc acid, decyl methylsulfoxide, urea, citric acid, Succininc acid, decyl methylsulfoxide, urea, 40 peanut oil, hemp, borage, olive oil, Sunflower oil, Soy peanut oil, hemp, borage, olive oil, Sunflower oil, Soy bean oil, monoi oil and macadamia oil; bean oil, monoi oil and macadamia oil; b. an active drug molecule selected from the group consist wherein said vasodilator and said dermal penetration ing of acetaminophen, acetohydoxamic acid, enhancer are covalently linked. acetophenazine, acyclovir, albuterol, allopurinol, 3. A bifunctional molecule comprising 45 amiloride, amoxicillin, amphetamine, amplicillin, a. a vasodilator selected from the group consisting of amrinone, arginine, bamethan Sulphate, bencyclane atenolol, baclofen, beclomethasone, betamethasone, fumarate, benfurodil hemisuccinate, benzyl nicotinate, budesonide, bumetanide, butorphanol, carbamazepine, buflomedil hydrochloride, buphenine hydrochloride, carphenazine, cefuroxime, cephradine, chlorampheni butalamine hydrochloride, cetiedil citrate, ciclonicate, col, chlorothiazide, chlorZoxaZone, cinoxacin, cloraZe cinepazide maleate, cyclandelate, di-isopropylammo 50 pate, cloxacillin, cyclacillin, dapsone, dicloxacillin, nium dichloroacetate, ethyl nicotinate, hepronicate, diethylstilbestrol, dopamine, doxorubicin, erythropoi hexyl nicotinate, ifenprodil tartrate, inositol nicotinate, etic, estradiol, fenoprofen, human growth hormone, isoXSuprine hydrochloride, kallidinogenase, methyl hydralazine, hydrochlorothiazide, ibuprofen, nicotinate, naftidrofuryl oxalate, nicametate citrate, nic indomethacin, insulin, isoproterenol, levodopa, levothy eritrol, nicoboxil, nicofuranose, nicotinyl alcohol, nic 55 roXine, meclofenamate, melphalan, metformin methyl otinyl alcohol tartrate, nitric oxide, nonivamide, Oxpen Salicylate, metronidazole, minoxidil, morphine, nad tifylline, papaverine, papaveroline, pentifylline, olol, nalidixic acid, naproxen, nomifensine, norfloxacin, peroxynitrite, pinacidil, pipratecol, propento?yltine, Oxaprozin, paramethasone, perphenazine, phenylpro raubasine, Suloctidil, teasuprine, thymoxamine hydro panolamine, probenecid, quinethaZone, ritodrine, Sco chloride, tocopherol nicotinate, tolazoline, Xanthinol 60 polamine, serotonin, terbutaline, terfenadine, tocainide, nicotinate, diaZoxide, hydralazine, minoxidil, sodium triamterine, trimethoprim, and Valacyclovir, nitroprusside, clonidine, quanaberZ, methyl dopa, wherein said dermal penetration enhancer and said indoramin, phenoxybenzamine, phentolamine, pra active drug molecule are covalently linked. Zosin, bedmidine, debrisoquine, guanethidine, k k k k k