Dec. 29, 1970 R. J. HERSCHER 3,551,554 ENHANCING TISSUE PENETRATION OF PHYSIOLOGICALLY ACTIVE AGENTS WITH DMSO Filed Aug. 16, 1968

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O 3Oar (262 /2O 24O INVENTOR. T/ME/W M/NL/7A5 Robert J. HerSchler d.14%. Attorneu 3,551,554 United States Patent Office Patented Dec. 29, 1970 1. 2 to pass freely, while rejecting others or permitting only 3,551,554 ENHANCING TISSUE PENETRATION OF PHYSIO slight passage. Such membranes comprise the body cover LOGICALLY ACTIVE AGENTS WITH DMSO ings and externally communicating cavities, including the Robert John Herschler, Camas, Wash., assignor to Crown skin and mucous membranes of the body cavities, e.g. Zellerbach Corporation, San Francisco, Calif., a cor alimentary tract, respiratory tract, genitourinary tract, poration of Nevada oral cavity, eyes, etc. (collectively defined herein as ex Continuation-in-part of application Ser. No. 329,151, ternal membranes). They also include internal mem Dec. 9, 1963. This application Aug. 16, 1968, Ser. branes such as the linings of the various organs and other No. 753,231 internal body structures, e.g. peritoneum and pleura, and Int, C. A61k 27/00 O the membranes surrounding cellular and intracellular U.S. C. 424-7 42 Clains structures. It is desirable in overcoming the aforemen tioned problems in drug administration, to increase the passage or penetration of agents across such membranes ABSTRACT OF THE DISCLOSURE and further to enhance their intercellular and intracellular A method of enhancing tissue penetration of phys 5 diffusion in order for them to reach their situs of activity iologically active agents, including physiologically active more rapidly to achieve the desired response more quick steroids, antineoplastic agents, antigens, anti-unicellular ly and often more effectively. It is exceptionally desirable microorganism agents, antihistaminic agents, neuro to do this in a reversible manner, by which is meant pene pharmacologic agents, antiinflammatory agents, anti tration of the agents into tissue without adversely affect coagulants, vasodilators, ultra-violet screening agents, di 20 ing or impairing the function or structure of the tissue. agnostic dyes and radiopaque agents and nutrients, by It is known that certain substances will penetrate tissue conjointly applying them to the tissue with dimethyl sulf only after the tissue has been irreversibly damaged which oxide. Penetration of the skin and the mucous mem is certainly undesirable. Certain agents, such as sur branes of the body cavities by these agents may be en factants, have been known previously for increasing pene hanced by conjoint application of such agents and di 25 tration of various agents. However, again, such pene methyl sulfoxide directly to such membranes. Preferably, tration was effected only through irreversible damage of for penetration of agents through the skin compositions the tissue. of DMSO at concentrations of 50% and above are em It has been a major rule in medicine that the “vehicles' ployed and for penetration through mucous membranes, or "carriers' have relatively little effect on the penetra compositions including DMSO at concentrations of 10% 30 tion rate for a given agent and this rule generally still and above are employed. Antineoplastic agents, steroids, holds true. Thus, with conventional carriers for medi central nervous system-active agents, local anaesthetics, cines, such as alcohol, carbowax, water, etc., few agents anti-inflammatory agents, diagnostic dyes and radiopaque will adequately penetrate such formidable external mem agents, and vasodilators may be advantageously adminis brane barriers as the intact skin or mucous membrane. tered by injection with DMSO in concentrations pref 35 It is to be expected that this would be true of all po erably up to 20% by weight to enhance penetration of tential "vehicles' or materials combined with physiologi internal tissue membrane barriers to achieve better distri cally active agents. However, surprisingly, it has been dis bution of these agents. covered that dimethyl sulfoxide (DMSO) has the unusual ability to greatly enhance the penetration of agents when 40 they are applied to such membrane barriers along with CROSS REFERENCES TO RELATED APPLICATION dimethyl sulfoxide. The penetration of agents which previously have not penetrated these membranes to an This is a continuation in part of co-pending application effective degree may be enhanced sufficiently so that a Ser. No. 329,151, filed Dec. 9, 1963, now abandoned. useful result may be obtained. The penetration of agents BACKGROUND OF THE INVENTION 45 which have been known to penetrate to a limited degree A predominant and limiting problem in the develop in conventional vehicles may be significantly enhanced. ment and use of physiologically active agents is the in New and convenient routes of administration, often with ability to administer them as effectively as is desired. In a decrease in side effects of the agents, better localized particular, there is often a limitation as to the routes of concentration and a more Sustained activity, may there administration because of the following factors: 50 by be created for many agents. (1) Some agents are inactivated in the gastrointestinal In my co-pending application (Ser. No. 615,377 filed tract or they are absorbed poorly into the body from the Feb. 13, 1967) is disclosed my related discovery that tract. Also, undersirable side effects may result which DMSO enhances the penetration of plant-active agents prevent effective oral administration. 55 (pesticides, dyes, nutrients, hormones, herbicides, and (2) In every case where injection must be restorted to, the like) into plant tissue in a highly unusual manner. there is a risk of needle injury, infection, and other trau Dimethylsulfoxide is a water-white liquid at room tem ma (including the emotional trauma inevitably associated perature having a freezing point of approximately 18.5 with injections). C. and a specific gravity of approximately 1.1. Dimethyl (3) Few agents are absorbed through the skin or sulfoxide is a well known industrial solvent and it has 60 been available in commercial quantities for at least a mucous membranes in effective quantities and the rate decade (from Crown Zellerbach Corporation, San Fran of absorption is less than would be desirable for those cisco, Calif.). DMSO was originally synthesized in 1866 that do. and since that time it has been extensively investigated (4) A local concentration for a local effect is often for possible industrial and biological utility and a con desired but a larger systemic dose must be given to 65 siderable amount of literature has developed on its prop achieve an effective concentration at the local area when erties and uses. Over the last 25 years it has found wide the agent can only be injected or given orally, (but spread use as a solvent in industry and in the laboratory. not topically). This higher dose often causes undesirable DMSO has been investigated in the past for various side effects, since dosage related side effects are very biochemical uses, for example as a reaction solvent for prevalent for many agents. 70 preparing derivatives of various proteins, and antibiotics, Animal tissues comprise various membranes which are as an extraction solvent for various proteins, as an selectively permeable and which allow some substances analytical solvent and as a solvent for various other lab 3,551,554 3 4. oratory uses. It has also been suggested as a solvent for systemic effect for all of the usual injectible routes, e.g. certain pesticides (see, for example, U.S. Pat. No. 3,068, subcutaneous, intramuscular, intraperitoneal, etc. 142). Where a local effect is desired, the intimate distribution Buso has been investigated as a preservative agent of the agent in the tissue near the site of injection pro for in vitro storage of chilled or frozen tissue and it has longs and enhances its physiologic activity at this local also been determined to have a protective effect in ex 5 site. This may permit use of a lower dose to achieve the perimental animals subjected to X-irradiation follow desired response with a smaller risk of side effects which ing injection of DMSO into such animals. may result from a higher dose. In connection with topical application of the antifungal For all routes of administration, conjoint application griseofulvin, DMSO has been listed along with various of DMSO along with physiologically active agents hav inert materials as “bland, high boiling fluids' to be used O ing an activity site in the individual cells of the host may as carriers for the griseofulvin in applying it to the skin additionally result in an enhanced effect of the agent to control fungus growth in the skin (see British Pat. No. through the ability of DMSO to increase the permeability 810,377). DMSO has been employed as a solvent for of Such individual cells to such agents. preparation of certain injectable formulations, namely As previously indicated, the mechanisms of penetration chloramphenicol and an anthelminic preparation (see Pat. enhancement are as yet not fully elucidated. Accordingly, Nos. 3,044,936 and 3,067,096). it is not intended to be bound to one specific theory of Despite the employment of DMSO as a solvent for operation. However, it is believed that DMSO acts by these purposes and despite general experimentation with Several mechanisms in enhancing penetration. DMSO is DMSO in the medical field, the unique ability of DMSO believed to act directly on tissue to alter the general per to alter membrane permeability and to thereby enhance meability of the tissue membrane. More specifically, penetration of physiologically active agents was neither DMSO when applied thereto, is believed to decrease the suggested nor discovered. Although DMSO has been a natural resistance of tissue membranes to penetration by well known and widely investigated solvent for many foreign agents. DMSO is also believed to promote pene years, its unique ability to enhance penetration of external tration by a direct transport effect, perhaps by the mecha and internal membrane barriers as contemplated in the nism of complexing with the agent. This mechanism is be present invention has been totally unrecognized. lieved more applicable to cationic and anionic agents. SUMMARY OF THE INVENTION GENERAL DESCRIPTION OF THE INVENTION By a mechanism or mechanisms not yet fully under This invention is applicable to the tissue or organisms stood, DMSO, when applied to animal tissue, increases of all animal phyla, DMSO having differing degrees of the permeability of the tissue in a reversible manner to influence on penetration of various tissue types of a given cause a much greater penetration rate for conjointly ap animal. Animals of particular importance in the practice plied physiologically active agents. Although the mode of the invention are the mammalians, especially man and of activity is still unclear, it is definitely not that of the veterinary animals. However, the invention may also be simple "vehicle' or “carrier' since the effect may be ob practiced with other vertebrates, as for example the tained to some extent even when the DMSO is applied amphibians, fishes, reptiles, etc. and with the lower species to the tissue separately and the enhanced penetrability of comprising the non-vertebrates. the tissue may last for as much as three hours after the As indicated previously, a measure of penetration en DMSO treatment. 40 hancement may be obtained where the tissue is pretreated When applied to the intact skin along with dimethyl with DMSO prior to application thereto of the physiologi sulfoxide, particularly at a DMSO concentration of 50% cally active agent. The tissue penetrability is thus altered by weight and above, or to skin pretreated with the di by Such pretreatment and this reversible effect gradually methyl sulfoxide, an agent such as a steroid, may pene diminishes and the tissue returns to its normal perme trate rapidly to and Saturate the stratum corneum (the ability State. However, for convenience and optimal effect, highly resistant "horny layer' of the skin which is the 45 it is frequently desirable to administer the DMSO and the major barrier to penetration). The steroid continues to agent simultaneously in the same composition. penetrate through the skin from this “reservoir' in the Penetration enhancement is generally non-selective in Stratum corneum to the underlying tissue and into the terms of the type or physiological effect or effects of circulatory system. agents to be transported across membrane barriers. The Similarly, penetration into underlying tissues and into extent of penetration enhancement will depend upon the circulatory System may be obtained from topical ap many factors, the predominant factors being the relative plication to the mucous membranes of the body cavities natural permeability of the particular membrane, the as in the case of intraoral, conjunctival sac, rectal, vaginal, concentration of DMSO applied, the extent of solubility and bladder instillation administration, particularly where of the agent in DMSO and the chemical and physical the DMSO is utilized at a concentration of 10% by weight 5 5 properties of the agent. and above. It is thus seen that a particularly important As a class, cationic agents, means chemical compounds aspect of this invention is that penetration of agents may Which dissociate into relatively small, mobile anion(s) be effectively enhanced following topical administration. and much less mobile cation(s) which are considerably As used in this connection herein, the term “topical is larger than the anion(s) (e.g. having a radical Weight intended to include application to all external membrane 60 ratio greater than 1 to 3, but more usually on the order barriers including the cutaneous or epidermis regions and of 1 to 10-100), appear to obtain the most pronounced the mucous membranes including the gastrointestinal penetration enhancement with DMSO. Even penetration tract, the respiratory tract and the genitourinary tract. of external membrane barriers such as the mucous mem Important advantages are also obtained through the in branes may be effected with these agents utilizing rather jection Toutes for physiologically active agents. When low amounts of DMSO, frequently as low as 10-20% by these agents are injected into the tissues either in a com Weight. Following Examples 34, 69, 73 and others illus position including dimethyl sulfoxide (preferably at trate cationic agents. Anionic agents, meaning agents DMSO concentrations exceeding 1% arid especially in the which dissociate into relatively small, mobile cation(s) range of 10-20% by weight) or together with conjoint, and large, less mobile anion(s) which are considerably but separate application of DMSO to the tissues, the effect larger than the cation(s) (e.g. having a radical weight is an enhanced and more even distribution thereof into ratio greater than three to 1 but more usually on the the tissues surrounding the injection site compared with Order of 10-100 to 1) also obtain marked penetration conventional injection techniques. This more even dis enhancement with DMSO. Although lower concentrations tribution is of considerable advantage for both local and of DMSO, as for example 15% by weight (see following 3,551,554 5 6 Example 34), may be effected through various external positions containing a DMSO concentration of between membrane barriers, higher concentrations of DMSO are about 50% and 90% by weight and containing water, frequently desirable for maximum effect, e.g. 50% by preferably 10% by weight or greater. weight and above, particularly for epidermal application. Application to mucous membranes follows generally These agents are illustrated by following Examples 34, 38, the procedure for cutaneous administration. However, 51, 70 and others. 5 lower concentrations of DMSO, for example as low as Penetration of non-dissociating chemical compounds 10% by weight, may be preferred since penetration of may also be beneficially enhanced with DMSO. Here mucous membrane is more easily affected. again higher concentrations of DMSO, i.e. 50% and above, For most injection routes preferably lower concentra are frequently desirable for maximum effect, particularly tions of DMSO of about 10% to about 20% by weight are for epidermal applications. Illustrative of this class of O preferably utilized. For some injection routes for example, chemical agents are following Examples 1, 2, 3, 4, 47, intra- and peri-articular routes, higher concentrations, say 58, 72 and others. 30-40%, may be preferred. Penetration of agents which form complexes with The amount of the physiologically active agent to be DMSO, for example iodine and most metal halides and administered will obviously be an effective amount for nitrates, are also beneficially enhanced. Such agents are the desired result expected therefrom. This, of course, will illustrated by Examples 41, 42, 43 and 74. be ascertained by the ordinary skill of the practitioner. The size of the compound obviously may influence to Due to enhanced activity which may be achieved through some extent the relative ability of agents to penetrate tis better penetration, the dosage of agent may often be de sue. However, effective membrane penetration utilizing 20 creased from that generally applicable. In accordance DMSO has been demonstrated for extremely large com with the usual prudent formulating practices, a dosage pounds, for example compounds having molecular weights near the lower end of the useful range of the particular exceeding 40,000 (also see Examples 75 and 76 illustrat agent may be employed initially and the dosage increased ing penetration enhancement of insulin which has a molec as indicated from the observed response, as in the routine ular weight of about 6,000). Even for such a formidable 25 procedure of the physician. membrane barrier as intact human skin, quite large com As previously discussed, the DMSO may advantageous pounds have been demonstrated to be effectively en ly be compounded with the physiologically active agent hanced. As illustrated by following Example 68, heparin, for concurrent administration. The usual pharmaceutical which has a molecular weight of 8,000 and above, can be compounding agents, diluents or carriers may be included effectively penetrated through the human epidermis in 30 in these compositions as desirable for the particular route SO CaSeS of administration and dosage form. The amount and type Standard occlusion techniques frequently may increase of diluent or carrier used should, of course, be consistent the percutaneous absorption of the larger molecules. In with the compatability of the agent in DMSO and the dil general, at least a limited degree of solubility of the agent uent. A cosolvent or other standard adjuvant, such as in DMSO is desirable to achieve maximum benefit of the a surfactant, may be called for to maintain the agent in present invention. Naturally, the practitioner will select solution or suspension at the desired concentration. Where agents, routes of administration and composition forms stability of the agent in the presence of DMSO at the de guided by these well-known principles. sired concentration is a problem, it may be desirable to The term “physiologically active' in describing the prepare the formulation immediately before administra agents contemplated herein is used in a broad sense to 40 tion or to administer the DMSO and the agent separately comprehend not only agents having a direct pharmacolog to the tissue. ical effect on the host but also those having an indirect In selecting the route of administration for a given or observable effect which is useful in the medical arts, agent, obviously the known toxicity, side effects and ef e.g. the coloring or opacifying of tissue for diagnostic pur fectiveness for a given route of administration should be poses, the screening of U.V. radiation from the tissues, 45 taken into account. For example due to skin irritation etc. Agents, penetration of which across membrane bar known to be caused by Some agents or due to poor pene riers, particularly external membranes, may be beneficial tration characteristics, some other route than dermal ap ly enhanced upon direct application include: physiologi plication may be the route or choice for such agents. cally active steroids, antineoplastic agents, antigens, anti Dosage forms for topical application may include solu unicellular microorganism agents, antihistaminic agents, tions (paints), nasal sprays, lotions, ointments (including neuropharmacologic agents, anti-inflammatory agents, an creams and gels), suppositories and the like. The solutions ticoagulants, vasodilators, ultra-violet screening agents, di and nasal sprays may simply comprise the agent dissolved agnostic dyes and radiopaque agents and nutrients. Agents, in DMSO, optionally with an amount of water, glycerine the penetration of which across internal membranes may or other diluent. For nasal sprays and other mucous mem be particularly benefited upon injection include antineo 5 5 brane applications isotonic saline may be preferable as a plastic agents, steroids, central nervous system-active diluent. The DMSO may be present in these forms in agents, local anaesthetics, anti-inflammatory agents, diag various concentrations, say from about 10% to about nostic dyes and radiopaque agents and vasodilators. 75% by weight or higher. The concentration of the DMSO applied to enhance Lotions and gels, ointments or creams, may contain the penetration may vary over wide limits. The concentra 60 usual ingredients to provide a base, as for example cetyl tion selected is desirably related to the route of adminis alcohol, an emulsifier such as lauryl sulfate and water. tration to be employed. For cutaneous application, com Another base may be formulated by combining equal positions including at least about 50% by weight DMSO weight amounts of stearic acid, cetyl alcohol, triethanol are preferable in that they have been found to increase amine and glycerol monostearate with water. Still other percutaneous penetration in a highly significant manner. 65 bases may utilize polyethylene glycols of different vis with DMSO concentrations closely approaching 100% cosities, depending upon the desired consistency. DMSO Maximum cutaneous penetration is generally attained may be added to the lotion or ointment base in varying (excluding the agent), but with concentrations much amounts as desired, generally up to around 50% by above 90% by weight the incremental increase in pene Weight. tration rate over that achieved at 90% often is relatively O A Suppository form may be made from a high viscosity small. On the other hand, above a 90% concentration of polyethylene glycol 4,000, water and DMSO, which may dimethyl sulfoxide the side effects of a burning sensation be present in an amount of about 20% by weight. and erythema increase significantly. Accordingly, for top The concentration of physiologically active agent in the ical use, it may be desirable, consistent with physical sta various dosage forms is, of course, commensurate with bility of the composition, to formulate the DMSO in com 75 that normally utilized for the particular agent in conven 3,551,554 8 tional formulations for effective results for the intended antly in nature in animal and plant fats. Certain steroids route. Both the amount of physiologically active agent are naturally produced in the body and they act as hor and the amount of DMSO will be influenced by the type mones to mediate and control many body functions. of effect desired. If a more localized effect is required, as These hormones have been isolated or produced syn for example, in treating a superficial infection with an thetically and used in replacement therapy for hormone antibacterial agent, lower amounts of physiologically ac deficiencies. Additionally and importantly, these steroids tive agents and lower concentrations of DMSO may be have been found to be highly useful drugs in the treat called for. Where deeper penetration is desired, as in the ment of a wide range of disease states not primarily due case of local anaesthesia, a higher concentration of to lack of hormones. In the recent past extensive research DMSO may be desirable to promote adequate penetration. effort has resulted in the synthesis of a vast number of Where general systemic concentration of an agent is de O new steroid hormone derivatives having biological ac sired for a topical preparation, generally higher concen tivity (in excess of 1,500 compounds) and the list is grow trations of DMSO are desirable and the amount of agent ing rapidly. Such derivatives usually contain modifying as, for example, a steroid, may be included in the compo groups linked to the steroid nucleus which influenced the sition sufficient to provide the blood level desired. activity of the steroids, as by modifying, prolonging, or The various pharmaceutical forms are desirably pro increasing its activity, increasing stability and/or modify vided in determined amounts, as in containers of a given ing its solubility characteristics. These modifying groups volume. These amounts may include 100% DMSO con usually comprise addition salts to influence solubility or centration containing the desired dose of the agent, or a side chain substitution on one or more reactive ring a lesser concentration of DMSO with a diluent and the 20 carbon atoms. The side chain may be a direct substitution physiologically active agent dose. Thus, for example, for a ring hydrogen, an ester formed at a reactive hy graduated ampules containing, say 5 cc. of 100% DMSO droxyl group or an ether group. Many of these steroid with the agent dissolved therein may be provided. The derivatives have a higher potency than the naturally oc practitioner need only open and dispense all or a de curing steroid hormones, often with a decrease in the termined part to a subject. Nasal spray bottles, aspirators, 25 undesirable side effects which frequently result from suppositories, cotton tipped stick applicators, Squeeze administration of the natural steroids. As used herein, the tubes may all be utilized for topical application. term "steroids' and "steroid drugs” is intended to com The following illustrates the practice of the present in prehend both the natural steroids and the biologically vention with the various classes of agents. active modifications, derivatives and equivalents. 30 Steroid drugs may have one or more of many types of DESCRIPTION OF PREFERRED EMBODIMENTS biological activity such as anabolic, androgenic, gluco Alteration of membrane permeability cortoicoid, mineralocorticoid, estrogenic, progestogenic, lipoidiatic (removal of stored fat), circulatory system ac The following example is an invitro demonstration of tivity, central nervous system activity, anti-cancer and the effect of DMSO on the penetrability of tissue mem anti-Osteoporic. Some steroid drugs have the ability to branes. block the activity of other hormones, including the activ EXAMPLE 1. ity of other steroids. Their biological activity may be char Penetration of solutes and ions through skin of frogs acterized as antiandrogenic, antiglucocorticoid, antimin The skin of the frog Rana pipiens is often used as a eralocorticoid (diuretic), antiestrogenic and antiprogesto “model membrane' system. The skin is removed and 40 genic. The various activities will be considered hereinafter placed as a wall barrier between two glass chambers in connection with specific embodiment of this invention. which are filled with dilute sodium chloride Salt solution. GLUCOCORTICOIDS The chambers are constructed to permit electrical measurements and addition or sampling of fluid from Hormones produced naturally in the body, by the adre either side. The fluid in the one chamber bathes the outside 45 nal cortex are called adrenal cortical steroids or corticoids. of the skin and the fluid in the other chamber bathes the Some corticoids, predominantly cortisone and hydrocorti inside of the skin. When water, irons or molecules cross Sone, have the property of influencing the rate of metabo from the inside to the outside solution, the movement is lism of glucose. Hence, they, along with their derivatives termed an “outflux.” A material that would make a mem and modifications, are called glucocorticoids. Glucocorti brane more permeable to any substance is said to increase 50 coids have other predominant physiologic activity which a flux. makes them highly useful as drugs. One of the most im Radioactive ions and radioactive labelled compounds portant activities is the suppression of inflammation, par were placed in the inside compartment and allowed to ticularly in the treatment of arthritis and rheumatic cross. The movement was quantitized by making Serial diseases. Glucocorticoids are also useful in the treatmen" radioassay of the bathing fluids. The outflux rate of a given 55 of dermatoses, drug reactions, bronchial asthma, lupus substance was determined during several control periods. erythematosus, angioneuroedema and many other dis Dimethyl sulfoxide was then added to a 2.5% concentra orders. Masive does of corticosteroids have also been tion and flux measurements were made for several suc used to induce remissions in leukemia. DMSO may be cessive periods. The following table presents sample data combined with glucorcorticoids to enhance their penetra tion to the affected tissue for these various disorders. The to illustrate flux rates. 60 following examples illustrate compositions and treatments for this purpose utilizing the most prominent and active Ed. 15 min. Molesii.5min, natural and modified glucocorticoids: Substance Na Cl- Thiourea. Sucroso EXAMPLE 2 Control flux------0.36 0.14 0.0026 0.0020 Flux after dimethylsulfoxide.----- 1.27 1.53 0.0152 0, 0084 65 Penetration of injected corticosteroids Multiple flux increase------3.5 6 4 A thirty-two year old white woman was seen with a two In a number of similar tests the range of multiple flux days history of left subdeltoid bursitis. This gave her increase for these ions and molecules was about 5 to 10. pain on minimal abduction particularly, but also was 70 present in other movements of the shoulder joints. Physi STEROIDS cal examination revealed marked tenderness to pressure Steroids are generally classed as organic molecules With obvious protective muscle spasm overlying the joint. which have in common a perhydrocyclopentanophenan Two ml. of hydrocortisone was injected into the bursal threne nucleus and they are so named because they are area. This was associated with three hours relief of pain. related to and usually derived from sterols found abund 75 The patient was seen again two days later. At this exam 8,551,554 10 ination, her pain was just as marked as the first visit. Two dispersed. The triethanolamine is then added dropwise to ml. of hydrocortisone was injected and 5 cc. of 100% the mixture until it has gelled, care being taken to mini dimethyl sulfoxide was applied liberally to the entire left mize the air entrapment. This gel is particularly effective shoulder area. Within 15 minutes all pain disappeared and in the treatment of seborrhea and other scalp and hair in the patient reported no return of her symptoms when ex flammatory conditions and may be applied in amount and amined one week later. frequency conventionally used for topical application of this steroid. Better penetration and thereby an increased EXAMPLE 3 anti-inflammatory active is obtained for the amount of Penetration of injected corticosteroids steroid applied than results from its application in con A forty-two year old white male was examined with a O ventional formulations. one week history of acute subdeltoid bursitis of the right shoulder. Four mg. of Decadron was injected into the right EXAMPLE 7 subdeltoid bursa. The patient estimated that about a 20% The following ointment formulation may be prepared relief of his discomfort was attained. Full pain returned containing about 0.1% to 1.0% prednisone and preferably one day later. At this point, he was reinjected with 4 mg. 5 0.5%: of Decadron and 4 cc. of 100% dimethyl sulfoxide was Prednisone ------gm-- 0.1-10 placed on the skin over the involved bursa. This time the Glyceryl monostearate, acid type ------Sn-- 180 pain dissappeared completely and did not recur. Stearyl alcohol ------rs are me a mm or sala------m. gll-- 50 Both of the above examples show that cortisone injected Polysorbate 80 ------CC.-- 20 for subdeltoid bursitis is obviously of benefit, but that its 20 relief of symptomatology is enhanced with the simul Water ------CC-- 450 taneous application of dimethylsulfoxide to the skin. Dimethyl sulfoxide ------CC-- 300 The product is prepared as described in Example 5. The EXAMPLE 4 ointment is a valuable base for application of the corti Penetration of corticosteroids costeroid to inflammatory dermatological areas, particu A twenty-four year old medical student was seen with larly when they require inunction. Application is in ac atopic dermatitis of the right antecubital fossa. Three cc. cordance with that usual for topical application of this of 100% dimethyl sulfoxide were applied four times daily steroid in conventional bases. for three days. No benefit was noted. One mg. or 4 cc. 30 EXAMPLE 8 of Decadron (dexamethasone 21-phosphate) was applied The following cream formulations may be prepared four times a day for two days without benefit. One mg. containing about 0.1% to 1%. 16a-methyl prednisolone of dexamethasone 21-phosphate in 3 cc. of 100% di and preferably 0.5%: methyl sulfoxide was painted onto the involved area four Gm. times daily for three days. At the end of this period all 16a-methyl prednisolone ------0.1-10 evidence of the inflammatory reaction had disappeared. Stearic acid ------200 This example shows an improved action of dexametha Glyceryl monostearate, acid type ------200 sone 21-phosphate when used with dimethyl sulfoxide. Sodium lauryl Sulfate ------20 EXAMPLE 5 Dimethyl sulfoxide ------200 40 Water qs. 1000 cc. The following lotion formulation may be prepared con taining about 0.01 to 1.0%, with preferably 0.1% fluo As above, the product is prepared as directed in Ex cinolone acetonide: ample 5 and is useful in severe dermatoses requiring in Gm. unction. Fluocinolone acetonide ------0.1-1.0 Mineralocorticoids Cetyl alcohol ------200 45 Some natural corticoids have the predominant property Propylene glycol ------100 of inducing sodium retention (depressing the rate of ex Sodium lauryl Sulfate ------15 cretion of sodium salts through the kidneys). Hence, they, DMSO ------300 along with their derivatives and modifications, are called Water q.s. 1000 cc. 50 mineralocorticoids. The principal natural mineralocorti The steroid is dissolved in the DMSO and added to a coid is aldosterone, while desoxycorticosterone is the most stirred, cooling melt of the other ingredients. The prepa prominent synthetic mineralocorticoid. Both are useful in ration is particularly useful for the treatment of inflammed treating the mineralocorticoid deficient state in Addison's dermatoses by topical application to the affected skin area. disease. The following exemplifies the use of DMSO in The amount and frequency of application is in accordance the administration of mineralocorticoids to enhance pene with standard practice for topical application of this 55 tration: steroid. Penetration of the steroid into the inflammed tis EXAMPLE 9 sue is enhanced and a therapeutic level is achieved more The following ointment formulation may be prepared rapidly than when the steroid is applied in conventional containing about 0.5% to 2.5%, preferably 1.0%, de formulations. 60 soxycorticosterone acetate. EXAMPLE 6 Desoxycorticosterone acetate ------gm-- 5-25 The following ointment (gel) formulation may be pre Stearic acid ------gm-- 300 pared containing about 0.2% to 1.0%, and preferably Cetylaclohol ------gm-- 100 0.6% triamcinilone acetonide: Polysorbate 20 ------cc-- 20 Gm. 65 Sorbital 70% ------cc -- 100 Triamcinilone acetonide ------0.2-10 Dimethyl Sulfoxide ------cc 300 Polyethylene glycol 400 ------400 Water, q.S. 1000 cc. Dimethyl sulfoxide ------59 The product is prepared as specified in Example 5. The Carboxy vinyl polymer powder ------1. 70 product may be employed in treatment of pigmentation Triethanolamine ------0.4 in Addison's disease by topical application to the affected The corticosteroid is dissolved in a mixture of the first area. Penetration may be increased sufficiently so that two ingredients, and the carboxy vinyl polymer gelling effective results may be obtained. In conventional bases agent is sprinkled on the surface of the combined liquids this steroid has had very limited effectiveness topically and and stirred until all the particles have been wetted and 75 injection usually must be resorted to. 3,551,554 11. 12 Androgens lows containing about 1% to 5%, preferably 2%, testo Androgen is the generic term which comprehends tetro sterone propionate: sterone, the natural male hormone, androsterone and the Gm. modifications and derivatives thereof which have mascu Testosterone propionate ------10-50 linizing activity, Natural and modified androgens are em 5 Dimethyl sulfoxide ------890 ployed in replacement therapy for hypogonadal males. Water ------100 Typical for this purpose are the isobutyrate, decanoate, The steroid is dissolved in a mixture of the dimethyl isocaproate, enathate, phenylproprionate and cyclopentyl sulfoxide and water. The formulation may be applied proprionate esters of testosterone and 17-methyl testo topically as an anabolic and in the treatment of breast SterOne. O cancer. The enhancement of penetration over that ob However, a much more important drug roll for these tained with conventional topical formulations permits steroids is their use as antiestrogens in treatment of female more effective topical use of this steroid which previously genital cancer and as anabolic agents (metabolism stimu had to be injected to achieve a response in many cases. lation) in triating deboilitated subjects. For these pur poses, androgens which have been modified to decrease EXAMPLE 14 their unwanted virilizing effects (while retaining their ana The following cream may be formulated with the fol bolic and antiestrogenic activity) are generally preferred. lowing composition containing about 1% to 10%, and The following steroids exemplify these compounds with preferably 3%, 2a-methyl-dihydrotestosterone propionate the commercial source of the compounds indicated: (metholone): 17a-ethyl-19-nortestosterone (Nilevar) 20 17a-methyl-19-nortestosterone (Syntex product) Metholone ------gm-- 10-100 19-nortestosterone phenylproprionate (Durabolin) Steary acid ------II.-- 200 9a-fluoro-11B-hydroxy-17a-methyltestosterone Glycerol monostearate, acid type ------Il-...- 200 4-chloro-19-nortestosterone acetate (Halotestin) Sodium lauryl Sulfate ------CC-- 20 4-hydroxy-17a-methyl testosterone (Oranabol) 2 5 Water ------CC-- 400 2-hydroxymethylene-17a-methyl-dihydrotestosterone Dimethyl Sulfoxide ------CC-- 200 (Adrod-Parke Davis) The cream is prepared as directed in Example 5. The 17a-methyl-17B-hydroxyandrostano (3,2-C)-pyrazole product is useful in the treatment of muscle wasting and (Androstanazole) 30 weakness followed breast cancer surgery and may be ap 1-dehydro-17a-methyltestosterone (Dianabol) plied topically to the affected area. Penetration of the The following examples illustrate the use of DMSO steroid is greatly improved over that obtained in conven to enhance the penetration of androgens: tional formulations. EXAMPLE 1.5 EXAMPLE 10 A suppository formulation may be prepared as follows The following ointment (gel) may be formulated con containing about 1 to 5%, preferably 2%, testosterone taining about 1% to 5%, preferably 2%, steroid: propionate: 2-hydroxymethylene-17a-methyl-dihydrotesto Sterone ------gm... 10-50 Testosterone propionate ------gm-- 10-50 4) Propylene glycol ------cc - 500 Polyethylene glycol 4000 ------gm-- 400 Dimethyl sulfoxide ------cc. 498 Propylene glycol monostearate ------gm-- 100 Carboxy vinyl polymer powder ------gll-- 1. Dimethyl Sulfoxide ------cc 500 Triethanolamine ------9IIl-- 0.5 The solid constituents are melted, added to the solu The product is prepared as specified in Example 6. The tion of the steroid in DMSO and poured into an appro product is useful in topical anabolic treatment, partic priate mold. The product is recommended for rectal appli 45 ularly in preventing thinning of the skin and in inducing cation as replacement therapy. blood vesel thickening. EXAMPLE 11. Estrogens and progestins A suppository formulation may be prepared as follows containing about 1 % to 5%, preferably 2%, 17-methyl Estrogen is the generic name for estradiol and its active tetrosterOne: metabolites estrone and estriol, naturally occurring fe male sex hormones, and their derivatives and modifica 17-methyl testosterone ------gm-- 10-50 tions. Estrogens are useful in treating menstruation dis Hydrogenated castor oil ------gm - 400 orders, infertility, habitual abortions, and endometriosis. Stearic acid ------gm-- 100 Along with progestrogens, they are used to control the Dimethyl Sulfoxide ------cc 500 reproductive cycle in women for contraception. They are The product is prepared as noted in Example 9 and also used in replacement therapy, particularly in treating used in a similar manner. the hormone deficiency states such as in postmenopausal EXAMPLE 12 Women. Modified estrogens having lower feminizing char 60 acteristics are particularly useful for other applications in A cream formulation may be prepared as follows con cluding the treatment of atherosclerosis and osteoporosis. taining about 1% to 10% preferably 3%, 17a-ethyl-19 Their antiandrogenic effects are also useful in the treat notestOSterOne: ment of prostatic cancer. 17a-ethyl-19-nortestosterone ------gm-- 10-100 Progesterone is a natural female hormone which plays Cetyl alcohol ------Sn-- 250 a primary role in the reproductive cycle of the female Stearyl alcohol ------g-- 200 mammal, particularly in the menstrual cycle of the pri PolySorbate 80 ------CC-- 20 mate. Progesterone and its modifications and derivatives Water ------CC-- 250 are classified as progestins. Progestins are useful in re Dimethyl Sulfoxide, q.S. ------cc. 1000 placement therapy and in treatment of menstrual dis This cream may be prepared as noted in Example 5. orders and prevention of fetal loss. A number of modified progestogens are useful in contraception. The various It may be applied topically for stimulation of epitheliza modified progestogens include the 19-morprogestogens, tion and connective tissue regeneration. the 17a-methyl progestogen derivatives, the 3-enol ethers EXAMPLE 13 of progesterone, and 17-acetoxyprogesterone and the 9 A lotion or paint formulation may be prepared as fol iso-10-iso compounds called retroprogesterones. 3,551,554 13 14 The following exemplifies the practice of this inven Spirolactones tion with estrogenic and progestenic steroids: Hypersecretion of aldosterone (primary aldosteronism) EXAMPLE 16 as a primary event or secondary to other disease states, The following lotion may be formulated as follows Such as cardiac, renal, and hepatic disorders (secondary containing about 0.1% to 1.0%, preferably 0.4%, estra aldosteronism) may cause undesirable salt and water re tention and promote edema. A certain steroid series, the diol valerate: 17-spirolactosteroids or spirolactones, possesses an anti Estradiol valerate ------gm-- 1-10 aldosterone activity capable of blocking the effects of Cetyl alcohol ------gm-- 200 aldosterone on the kidney. Hence, they are called aldo Propylene glycol ------gm-- 100 IO sterone antagonists. The following example describes use Sodium lauryl Sulfate ------gm-- 15 of the spirolactones: Water ------cc. - 400 Dimethylsulfoxide ------cc. 300 EXAMPLE 21 This product is prepared as noted in Example 5. The The following ophthalmic formulation may be prepared product is designed as a means of establishing systemic containing about 0.1% to 0.75%, preferably 0.3%, spiro replacement therapy for estrogens during menopause by nolactone: simple topical application to the skin or mucous mem Spironolactone ------gm-- 1-7.5 brane. The DMSO enhances penetration of the estrogen Polyethylene glycol 4000 ------cc - 200 sufficiently to obtain a systemic effect. This has not been 20 Dimethyl Sulfoxide ------cc. 200 possible in conventional formulations. Water, q.S. 1000 cc. EXAMPLE 17 The formulation is prepared by melting the polyethyl A suppository may be formulated as follows to contain ene glycol 4000, dissolving the steroid in the DMSO, mix 0.1 to 1.0%, preferably 0.5%, of 3-methyl ether of eth ing the two liquids together and diluting to volume with eynylestradiol: 25 water while stirring. The preparation is applied topically 3-methyl ether of ethynylestradiol ------gm-- 10-100 to the eye by eye dropper, or similar applicator, for treat Polyethylene glycol 4000 ------girl-- 400 ment of glaucoma. Propylene glycol monostearate ------9Il-- 00 Antineoplastic agents Dimethyl sulfoxide (DMSO) ------CC-- 500 30 Antineoplastic chemical agents are drugs that combat The suppositories are prepared as noted in Example 11. cancerous processes. Antineoplastic agents at their pres The product is used in estrogenic replacement therapy ent stage of development are generally only palliative, for and may be used by rectal or vaginal application. in some instances only a temporary alleviation of subjec tive symptoms is obtained while the malignant process it EXAMPLE 18 self advances steadily. Nevertheless, these drugs play an The following ointment (gel) may be formulated con important role in the treatment of malignancies to induce taining 0.1% diethylstilbesterol: remissions. Diethylstilbesterol ------gm-- 1 A vast number of compounds have been screened for Propylene glycol ------cc - 500 40 possible anticaner action and screening programs are Dimethyl sulfoxide ------cc.-- 498 continuing in an attempt to develop safer and more effi Carboxy vinyl polymer powder ------gm-- 1 cacious antineoplastic agents. Of the screened compounds, Triethanolamine ------gm-- 0.5 only a fraction have proved of clinical experimental interest and a considerably smaller number have survived This gel is prepared as detailed in Example 6. The in practice. Many showing early promise have been preparation is particularly suitable for topical application 45 discarded because at useful dosage levels they have proved in the treatment of adolescent acne. too toxic for practical employment. Others have had only a low rate of successful results, due to the lack of ability EXAMPLE 19 to maintain an effective concentration at the site or to A cream may be formulated as follows to contain penetrate effectively into the neoplastic tissue and into about 0.72% norethynodrel and about 0.028.6% mestra 50 individual neoplastic cells. nol: These same problems also pose as a serious limitation on the effectiveness of those agents which have proved Norethynodrel ------gm-- 10.5 to be clinically useful. Specifically, in order to reduce and Mestranol ------gm-- 0.42 perhaps eliminate neoplastic cells, the antitumor agent Cetyl alcohol ------gm-- 100 55 must reach these cells in sufficient concentration. This Stearyl alcohol------gm-- 100 may be hindered or prevented by certain pharmacologi Polysorbate 80 ------cc.- 20 cal "barriers' or membranes. For example, in the case Water ------cc.-- 250 of leukemia, the blood-brain barrier, is an important ob Dimethyl sulfoxide, q.S. ------cc. 1000 stacle to effective treatment with many agents. In most in 60 stances the effectiveness of antineoplastic agents is also This cream is prepared as noted in Example 5. This ultimately limited because strains of neoplastic cells formulation is to be used as a contraceptive agent applied eventually develop which are resistant to their action. cutaneously twice monthly at a dosage of 10 grams. The survival of these cells is believed, in at least some in EXAMPLE 20 stances, to be the result of their natural resistance to the 65 entry of the antineoplastic agent. The proliferation of A suppository formulation may be prepared as follows: these resistant cells eventually causes the destruction of Chlormadinone ------mg-- 5 the host. Stilbesterol ------mg-- 1 With conventional carriers for medicines, such as Polyethylene glycol 4000 ------gm-- 400 alcohol, peanut oil, carbowax, etc., few antineoplastic Propylene glycol monostearate ------gm-- 100 agents will penetrate such formidable external membrane barriers as the intact skin or muceous membrane to effec Dimethyl Sulfoxide ------cc - 500 tively reach the neoplastic tissue. Also, there is little or The suppositories are formed as in Example 11. The no evidence that conventional carriers have any ability product may be employed for treatment of irregular or to alter the permeability of neoplastic tissue or to effec prolonged bleeding. tively enhance penetration of antineoplastic agents into 3,551,554 15 16 such tissue. Nor do such carriers facilitate penetration of body weight by inclusion in a solution and then direct in these agents into neoplastic cells. stallation in the freshly evacuated bladder of a dog. However, the penetration of antineoplastic agents which The two solutions were: previously have not penetrated certain membranes to an effective degree may be enhanced sufficiently with DMSO Solution 1 So that a useful result may be obtained through applica Thio-TEPA ------mg-- 5 tion to such membranes. Penetration of agents which have Isotonic Saline, q.S. ------ml -- 100 been known to penetrate to a limited degree in conven Trisodium phosphate ------M 0.05 tional agents may be significantly enhanced. Better local Phosphoric acid ------M 0.03 ized distribution, an increased activity and new and con O Dioctyl sodium sulphoSuccinate ------mg-- 0.5 venient routes of administration, often with a decrease Solution 2 in side effects of the agent may thereby be created for many antineoplastic agents. Additionally, DMSO has Thio-TEPA ------g-- 5 the ability by some mechanism not yet understood, to Isotonic Saline, q.S. ------mill 100 overcome the natural resistance of at least some malig Trisodium phosphate ------M 0.05 nant cells to penetration, thereby enhancing penetration of 5 Phosphoric acid ------M 0.03 antineoplastic agents into such cells to destroy them. Dioctyl Sodium Sulphosuccinate ------mg-- 0.5 Penetration into underlying tissues and into the cir Dimethyl sulfoxide ------w/v. 15% culatory system may be obtained from topical applica Blood counts were made periodically after installation. tion to the skin and mucous membranes. This is partic 20 The white count dropped approximately in half by the ularly useful for treatment of superficially located sixth day following administration of the composition tumors. Also, in treatment of localized tumors by the in containing dimethyl sulfoxide and the chemotherapeutic jection routes, improved localized distribution of the agent. The white count did not substantially drop in the antineoplastic agent in the tissue near the site of injec composition containing the chemotherapeutic agent tion by the use of DMSO may prolong and enhance its alone. physiologic activity at this local site. This may permit EXAMPLE 23 using a lower dose of agent to achieve the desired A lotion formulation may be prepared containing ap response with a smaller risk of side effect which may re proximately 10% cyclophosphamide and 80% DMSO sult from a higher dose. by blending the following: The same phenomenon may be of benefit for better 30 penetration of affected tissue in oral administration, in Grams regional therapy by intravenous and intraarterial infusion Cyclophosphamide ------10 and injection of antineoplastic agents and in perfusion Glycerine ------as a a a same a so a vers vs as use at - a sers a as as 10 techniques, such as those following surgical excision of CarboWax 4000 ------O tunOIS. DMSO ------80 Ultimately, for all routes of administration, this en This composition may be applied topically to the skin hancement of penetration increases the ability of the anti or mucous membrane adjacent the site of a localized neoplastic agents to cross membrane barriers which im cancer, or the like, typically at a dosage of 6-10 grams. pede their effectiveness to come in contact with malignant Antimetabolites cells in an effective concentration and to penetrate 40 malignant cells naturally resistant to penetration. Antimetabolites are generally analogues of naturally The following illustrates the practice of the present in occurring substances in the body necessary to cell metab vention with the various classes of antineoplastic agents olism. These agents destroy cancer cells by interfering when the classification "non-estrogenic anti-neoplastic with their metabolism. Since antimetabolites tend also to agents' is utilized, it is intended to mean all of the classes affect metabolism of rapidly multiplying normal cells, of antineoplastic agents, whether hormonal or non gastrointenstinal upsets and hematopoietic malfunctions hormonal, except for estrogenic anti-neoplastic agents, as as side effects limit the use and routes of administration for example esterone. of these agents. Regional treatment of affected sites is de sirable where effective. Principal antimetabolites are folic Alkylating agents acid antagonists such as amethopterin (methotrexate); Alkylating agents are highly reactive cyclic or unsatu 50 the purine analogues such as 6-mercaptopurine, azathio rated organic compounds having a denaturing or inactivat prine, 6-thioguanine and 8-azaguanine; and the pyrimi ing action on the malignant cell nucleus. A serious side dine analogues such as the uracils (principally the 5 effect of this group of agents is their tendency to damage halogen uracils, and 6-azauracil), azauridine and the 5 normal cells of the body. This causes a depression of halogen deoxyuridines. the blood-forming action of the bone marrow and injury 55 The following illustrates practices of this invention in to the gastrointestinal mucosa. This limits the type and connection with antimetabolic agents: routes of administration of these agents. Many are even too toxic to permit oral administration and hence regional EXAMPLE 24 treatment of the affected site is desirable where it can Methotrexate is a known folic acid antagonist used to be done effectively. 60 treat malignancy. Solutions containing this agent were The major types of alkylating agents are the nitrogen prepared as follows: mustards, the ethylenimines and the alkyl sulfonates. The Solution No. 1 more prominent agents include mechlorethamine, phenyl Methotrexate ------mg-- 5 alanine mustard, triethylene melamine (TEM), TEPA, Isotonic Saline ------cc - 50 thio-TEPA (tris (1-aziridinyl) phosphine sulfide), chlor 05 Evans blue dye ------mg-- 100 ambucil, busulfan, CRI-3, (a phosphinic amide mustard) Trisodium phosphate ------M-- 0:5 cyclophosphamide and mannitol mustard. These agents Phosphoric acid ------M-- 0.3 are employed in the treatment of Hodgkins' disease, vari Solution No. 2 ous forms of leukemia, metastatic cancer, various carci nomas and sarcomas. 70 Methotrexate ------I.-- 5 The following examples illustrate the practice of this Isotonic Saline ------ml. 42.5 invention in connection with alkylating agents: Evans blue dye ------mg-- 100 Trisodium phosphate ------M. .05 EXAMPLE 22 Phosphoric acid ------M. .03 Thio-TEPA was administered in a dose of 0.5 mg. per 75 Dimethyl sulfoxide ------ml - 7.5 3,551,554 17 18 Each of the solutions in an amount of 75 cc. was intro The melted blend is poured into a suppository mold to duced into the freshly evacuated bladders of dogs of about provide 10 gm. suppositories for treatment of prostatic equal size. Methotrexate is expected to pass the mucosa CaCe and basement membrane of the bladder to enter the vascu lar system, whereupon it is transported to the bone mar Antigens row where it will reduce the platelets in whole blood. A blood sample taken from the dog treated with solution Antigens are proteins, protein-polysaccharide com No. 2 five to six days after instillation, showed a w.b.c. plexes and polysaccharide which, when foreign to the drop from 600,000 to 100,000 per cc. This was the only blood stream of an animal and upon gaining access to the test solution which produced a physiological response. tissues of such animal, stimulate the formulation of a spe This illustrates the use of DMSO in achieving ade O cific antibody. These antibodies, synthesized by the animal quate blood levels of methotrexate for treating leukemia, in response to the antigenic stimulus, react with the in etc. This formulation may also be utilized for direct in vading antigen to render it harmless. However, despite this fusion into the arterial blood supply for localized neo defensive mechanism, antigens can be toxic when they plasms or for direct injection into accessible localized gain access to animal tissue, either through contact from neoplasms. exogenous sources or when produced in the tissue due to EXAMPLE 25 microorganism infection. As a means of diagnosis of antigen sensitivity and as A 5-fluorouracil formulation may be prepared by blend a prophylactic or desensitizing treatment to build up an ing the following: antibody defense against the invasion of antigens, such Grams 20 antigens, in specially prepared forms, are administered to 5-fluorouracil ------10 susceptible subjects. DMSO ------80 For example, in diagnosing allergy states, various sus Water ------10 pected antigens to which the subject may be allergic are Carboxymethyl cellulose ------0.25 applied intradermally or topically to the scratched skin The formulation is particularly useful in topical treat 25 of the subject. Allergy to a particular antigen is detected ment of skin tumors or other localized superficial tumors. from the characteristic wheal and flare response. In treat A typical dose is 5 grams. It is also useful in treating viral ing allergic states, desensitization may be achieved by in disorders such as warts. jection of an extract of the causative antigen in a series Vinca alkaloids 30 of gradually increasing doses. A more recent class of antineoplastic agents are the In prophylactic treatment to immunize subjects to the alkaloids extracted from the shrub Vinca rosea and their invasion of antigens of the infective type, toxoids or vac derivatives. The more prominent of these are vinblastine, cines are administered to the subject, usually by injection. winleurosine, vinrosidine and vincristine. The mechanism The antigenic effect of the toxoids and vaccines produces of action of the vinca alkaloids is still not known but they 35 specific antibodies which will then destroy subsequently have activity against a wide variety of neoplasms. invading antigens of specific types. The following examples illustrate the practice of this For these purposes, antigens generally must be ad invention in connection with vinca alkaloids: ministered by injection, because, in conventional formula EXAMPLE 26 tions, they are usually not effectively absorbed through 40 the intact or even abraded skin or mucous membrane. The Vinblastine sulfate may be administered I.P. to mice discomfort, risk of injury and infection and cumbersome at a dosage of 0.15 mg./kg. and concomitantly DMSO is procedures involved in such routes of administration are administered I.P. in 40% isotonic solution to a dosage of readily apparent. 2 gm./kg. Antigens may be administered topically with DMSO This example illustrates the specific use of DMSO, ad to the skin or mucous membrane to avoid the disadvan ministered separately, to enhance penetration of the anti 45 tages inherent in subdermal application. Additionally, the neoplastic agent from the general circulation into the neo DMSO, when combined with the antigen frequently has plastic tissue, a denaturing effect upon the antigen to decrease its toxicity Hormones to a more acceptable level. This may permit the avoid Various hormones, principally steroid hormones and ance of some of the usual extensive procedure required to their hormonally active substitutes, have activity against 50 treat the antigen to detoxify or attenuate it. Where cuta certain types of neoplastic conditions. The corticosteroids, neous application of antigens of a higher molecular as for example cortisone, hydrocortisone, dexamethasone weight is desired, they are preferably applied by occlu and prednisone are used in cases of lymphosarcoma, Hodg sion techniques and desirably they are applied to the more kin's disease, acute leukemia and mammary cancer me easily penetrated areas such as the axillary region. How tastases to obtain at least temporary remissions. Estro 55 ever, better results may be obtained with larger proteins genic compounds, as for example diethylstilbestrol, es and protein-polysaccharides by application to mucous trone, ethinyl, estradiol, dienestrol, chlorotrianisene and membranes. estradiol propionate, are used primarily in treatment of The following illustrates the practice of the present in prostatic carcinoma and to some extent in carcinoma of vention with various classes of antigens. the breast, chorioepithelioma and bladder cancer. Andro 60 genic steroids, principally testosterone, are employed Allergens primarily in treatment of breast cancer. For specific examples illustrating the practice of this Certain antigens cause an adverse reaction of the tis invention in connection with hormonal antineoplastic Sues of certain subjects, on exposure thereto (mediated agents, reference may be made to previous Example 13 65 by specific antibody formation), which antigens, in similar amounts, are innocuous to other persons. The condition and the following example: of sensitivity to such antigens is termed allergy and, ac EXAMPLE 27 cordingly, such antigens are also termed allergens. The A suppository formulation may be prepared by metling chief manifestations of the allergic reaction to allergens together the following: O are "hay fever,” asthma, gastrointestinal disturbance, Gm. urticaria, angioneurotic edema and serum sickness. Estradiol Valerate ------2.75 There are a wide variety of allergens and their mode Guiacol glycerol Stearate ------100 of contact is varied. These include such material as vege DMSO ------300 table and animal epithelial emanations, various pollens, Diglycol laurate ------150 75 such as tree pollens, grass pollen, pollen from the rag 3,551,554 19 20 weed family, and fungus spores, such as wheat and corn administered by injection in a solution or suspension. Ex rust and wheat Smuts. amples of conventional toxoids are tetanus and diptheria. As a diagnostic procedure to determine causative al toxoids. The various vaccines include those for hoof and lergens, a series of suspected allergens are screened by mouth disease, tuberculosis, cholera, influenza, typhoid contacting the tissue of the subject manifesting allergy fever, yellow fever, mumps, measles, whooping cough, symptoms with small amounts of the Suspected allergens. 5 and catarrhal (autogenous vaccines in mycotic infec The patient is then observed for a wheal and flare response tions). at the site of administration. A positive response is indica In this invention, DMSO is utilized to enhance penetra tive of allergy to the tested allergen. An extract of the tion of the conventional attenuated antigens. Additionally, confirmed allergen is conventionally prepared by Solvent DMSO may be used to assist in attenuating the antigens. extraction from extraneous matter with which it may be O The following examples illustrate compositions and im associated. The extracted antigen is then detoxified munization treatments utilizing a variety of infective anti through attenuation by heat and/or dilution with the genS: solvent. The denatured allergen is then administered to EXAMPLE 30 the subject in a series of increasing doses to desensitize the patient. In this invention, DMSO is utilized to en Immunizing formulations utilizing DMSO for enhanc hance penetration of these conventional attenuated ex ing penetration may be prepared utilizing the following: tractS. calf lymph smallpox vaccine The following examples illustrate compositions and tetanus toxoid, formaldehyde detoxified treatments, both diagnostic and desensitizing (hypoSensi killed typhoid and paratyphoid A-I-B bacillus tizing), utilizing a variety of allergens: Single immunization doses of each of these vaccines EXAMPLE 28 are prepared in a mixture of 0.5 ml. glycerine and 0.5 A twenty-four year old male subject had a four year ml. DMSO. 0.25% hexachlorophene is added as a pre history of recurrent hay fever supposedly based on an servative. allergy to tree pollenantigens. He was topically admin These formulations may be applied topically to the istered an allergenic extract of tree pollen on the upper mucous membranes, for example they may be applied as area of the left forearm. An inch below this area, the nose drops to the nasal passages. Application cutaneously extracted allergen in one co. of 100% dimethyl sulfoxide by air jet injector may also be advantageous. was placed. Below this area was placed one cc. of 100% dimethyl sulfoxide. After on hour only the composition 30 EXAMPLE 31 containing allergenic extract and dimethyl sulfoxide An influenza virus vaccine may be prepared by com showed a positive reading. The reading was evaluated as bining the following formaldehyde-inactivated virus a "marked' positive reaction. strains in a mixture of 7 ml. DMSO, 1 ml. glycerine and This same subject was later administered increasing 2 ml. water: doses of diluted tree pollen extract starting with an initial Units dose of 0.1 cc. placed into one cc. of 100% dimethyl Taiwan 1/64 (A2) ------1000 sulfoxide every seven days. The tree pollen allergenic dose Japan 170/62 (A2) ------1000 was increased but was always given with one co, of 100% FP 8 (A) ------'-- a------1000 dimethyl sulfoxide. The composition was applied directly 40 Ann Arbor 1/57 (A1) ------1000 onto the skin without resorting to injection by the usual Maryland 1/59 (B) ------2000 intracutaneous and subcutaneous route. The allergent An adult dose of 1-2 ml. of this formulation may be ad passed through the skin without a needle as shown by ministered twice at a six week interval, either cutaneously a characteristic reaction which occurred about three hours with an air jet injector or intranasally as nose drops. after each application. There was a marked diminution in the subject's symptomatology after three months of 45 EXAMPLE 32 treatment. A combination of types 1, 2 and 3 killed poliomyelitis EXAMPLE 29 virus strains in a concentration sufficient for one im Solution. A containing 2% of histamine in water was munizing dose is prepared in 1 ml. of: applied to the underside of the forearm. A like concentra Percent tion of histamine in 98% dimethyl sulfoxide containing DMSO ------80 2% water was also applied to the surface of the underside Ethanol ------10 of the forearm. A typical wheal was formed from the Water ------10 composition containing dimethyl sulfoxide, whereas no and protected by addition of 50 mcg. of neomycin base wheal was produced with the control histamine solution. 5 5 per ml. This shows that DMSO enhances histamine penetration. This formulation provides an adult dose which may While not, itself, an allergen, histamine is useful in hypo be applied intranasally as nose drops or as a spray. sensitization therapy in treating allergies. Cutaneous application by air jet injector may also be de Infective antigens sired. Serial doses are given in accordance with standard In a wide variety of infective diseases, prophylactic 60 procedure. treatment through immunization may be effective. Con ventionally, an infective microorganism is administered Anti-unicellular microorganism agents to the subject to be immunized in order to stimulate the DMSO is useful in enhancing the penetration and effec production of specific antibodies in the subject as a de tiveness of antimicrobial agents having growth-inhibiting fense against subsequent invasion of the infective micro 65 properties relative to unicellular microorganisms, i.e., organisms. Usually it is necessary to attenuate the micro anti-unicellular microorganism agents. By “growth-in organism preliminarily by denaturing it with heat or hibiting” properties it is intended to mean the exhibition chemicals or through the attenuating action of succes of biocidal or biostatic effects toward unicellular micro sive incubations or infective stages in living tissue. Some organisms. The phrase "unicellular microorganism' is in infective antigens, such as cowpox, utilized as an antigen 70 tended to include those microorganisms commonly desig for Smallpox, produce a low grade infection which pro nated by microbiologists as consisting of single cells, such duces antibodies specific against a more virulent micro as bacteria, viruses, rickettsiae, yeasts and protoza, as organism, e.g. smallpox. opposed to multicellular microorganisms such as most The vaccine or toxoid thus produced is conventionally 5 fungi DMSO provides enhanced penetration of the anti 3,551,554 21 22 microbial agent to the locus of the unicellular microor Solution was instilled into the urinary bladders of one of ganism. DMSO may additionally provide a lowering of the the dogs. resistance of the microorganisms to the anti-unicellular Aminophyllin in 10 gmc./kg. of body weight was microorganism agent, perhaps through action upon the inserted into the urinary bladders of the remaining dogs. cell wall of the microorganism. In one of these 15% dimethyl sulfoxide was added and There are many infections in animals caused by bac 5 in the other 15% normal saline. The sodium phosphate teria, and there are a good number of agents employed buffer was used to bring the volume of both the dimethyl to combat such bacterial action. Following is a discussion Sulfoxide and saline solutions to 75 cc. with a pH of 9. of Some of the more common antibacterial agents and All five test solutions were allowed to remain in the examples of the use of DMSO to enhance penetration of IO bladder for four hours. Blood samples were taken from these agents. W the femoral veins prior to instillation and at 30, 60, 120 The treatment of tuberculosis caused by the tubercle and 240 minutes after instillation. With respect to the first bacillus Mycobacterium tuberculosis is commonly effected three dogs, serum sulfadiazine and salicylate levels were With such drugs as amino salicyclic acid derivatives, iso determined on each sample as the diazotized derivative niazid, viomycin, dapsone, and pyrazinamide. The use of and the ferric salt respectively. Lee and White three-tube DMSO with these or other antituberculosis drugs may clotting times were recorded at the same intervals. Gross be effective in enhancing the penetration of these drugs to post mortem inspection was made to detect the presence the situs of the tubercle bacillus. Additionally, it may of the dye. render the bacillus more susceptible to the action of the Referring to the drawings, as seen in FIG. 1, clotting anti-tuberculosis drug. 20 time appeared significantly increased in the dimethyl sul foxide animals as compared to the saline control animals. EXAMPLE 33 As seen in FIG. 2, a six- to eight-fold increase of serum The following lotion may be formulated, containing Sulfadiazine was noted in the dimethyl sulfoxide animals about 13% by weight isoniazid: beginning at the initial 30-minute period over either the Gms. acetone or saline controls, Dimethyl sulfoxide ------80 As seen in FIG. 3, a two- to four-fold increase of serum Ethanol ------a a was a win - wers - a as as a' . . . . 10 Salicylate was present in the dimethyl sulfoxide dogs over Water ------'er------10 either the acetone or saline controls paralleling the serum soniazid ------15 Sulfadiazine rise. 30 Direct inspection of the urinary bladders at the con The above lotion may be employed in the treatment of clusion of the procedure revealed that in both the acetone tuberculosis verrucosis cutis by cutaneous applications of and Saline control animals, the Evans blue dye had pene 1 to 2 ml. to the situs three times daily. A saran-coated trated through the mucosa into the muscular layers of the occlusive bandage over the site of application may im bladder and on opening the peritoneal cavity, the bladder prove percutaneous absorption. had a faintly bluish tinge. In the dimethyl sulfoxide ani Urinary tract infections are caused by a variety of mals, however, the entire bladder was blue, and in addi bacterial organisms, and these may be treated with syn tion, the anterior peritonium overlying the bladder, the thetic chemotherapeutic agents such as hexamethylene retroperitoneal tissues and the contigous small and large tetramine, orthophenyl phenol and sulfonamides, and bowel had bluish tinges. broad spectrum antibiotics such as the tetracyclines and 40 As to the last two dogs, there was a two-fold increase penicillin. Again, the treatment of such infections with a in the absorption of aminophyllin expressed as milligrams combination of the usual chemotherapeutic agent plus per 100 cc. at 60 minutes after instillation into the bladder. DMSO may potentiate the action of the chemotherapeutic The dimethyl sulfoxide animal had 4.6 milligrams per agent upon the bacteria causing the infection by render cent while the saline control was 2.17 milligrams percent ing the microorganisms less resistant to the action of the 45 at the end of 120 minutes. After 240 minutes, the figures agent in addition to enhancing the penetration of the agent was respectively 5.75 milligrams percent for the dimethyl to the situs of the microorganism as in the case of a Sulfoxide dog and 3.77 milligrams percent for the saline urinary tract wash. control. The following examples illustrate a procedure useful for EXAMPLE 35 treatment of urinary tract disorders (as well as other 50 The following irrigation solution may be formulated disorders pertinent to the agents employed): containing about 5% by weight of hexamethylene EXAMPLE 34 tetramine: Gms. Five female dogs weighing between 11 and 13 kgs. Dimethylsulfoxide ------15 were anaesthetized with sodium pentobarbital. Each ani 55 Water ------80 mal was cathetized, its bladder emptied and the test solu Sulfisoxazole ------5 tion instilled through the catheter. The various test solu tions were made from the following basic solutions. The The above formulation may be employed as a twice gram percent and milligram percent figures relate to the daily instillation of 10 to 20 ml. for the treatment of number of such weight units per 100 cc. of a final liquid 60 urethritus. volume. EXAMPLE 36 Sodium Salicylate ------gm. percent.-- 5 The following antiseptic jelly may be formulated: Sodium sulfadiazine ------do---- 5 Gms. Evans blue dye ------mg. percent 62/2 Dimethyl sulfoxide ------10. Sodium heparin (145 units per mg.) ------do---- 125 65 Water ------70 Ortho phenyl phenol ------0.15 Three solutions were prepared from the basic solu Phenyl mercuric acetate ----.aer ------0.01 tions by adding 15% v./v. of dimethyl sulfoxide to one; Sodium carboxymethylcellulose ------1. 15% v./v. acetone to the second; and 15% v./v. of isotonic saline to the third. O The foregoing jelly may be packed in 3-gram plastic A sodium phosphate buffer system was added which tubes with a urethral nozzle for treatment of urethritus. was made from 0.05 M tribasic sodium phosphate, 0.33 Leprosy, like tuberculosis, is caused by an acid-fast M. phosphoric acid, and 0.67 sodium chloride, to make microorganism, Mycobacterium leprae. Leprosy leads to a total of 75 cc. with a pH of 9. deformities and paralysis from involvement of the nerves With respect to each of the solutions, 75 cc. of such 75 and from injuries of anaesthetic areas of the skin. Various 3,551,554 23 24 drugs are employed to treat this disease such as diamino diabetes, had a culture performed on his toe drainage. diphenyl sulfone (dapsone), chaulmoogra oil, and diethyl The findings were: Staphylococcus, coagulase positive, sen dithiol isophthalate (etusil). Treatment with a leprosy sitive to chloromycetin and erythromycetin, resistant to drug plus DMSO has led to some improvement in patients penicillin and tetracycline. suffering from this disease. An example of a suitable Application was made topically with 50% aqueous di formulation is illustrated in the following example: 5 methyl sulfoxide every four hours for 24 hours, and then a reculture of the toe drainage was made. It showed: EXAMPLE 37 Staphylococcus, coagulase positive, sensitive also to pen The following solution may be formulated containing icillin and tetracycline. dapsone as the active ingredient: Gms. O EXAMPLE 41 Dimethyl sulfoxide ------80 A 23-year old male with bilaterally infected ingrown Water ------m - - - 20 toenails on the great toe was treated on the right side with Dapsone ------1.5 a composition containing 2% iodine, 2.3% sodium iodide Morning and evening cutaneous application of the fore 5 and 95.7% dimethyl sulfoxide. The left great toe was going solution may be made directly to skin lesions. treated with a tincture of iodine. A single treatment for Penicillin is the drug of first choice in infections caused two minutes was employed. At the end of four days the by pneumococcus, streptococcus, gonococcus, Susceptible dimethyl sulfoxide-iodine composition reduced the infec strains of Staphylococci, Treponema, Clostridia, B. tion so that the right great toe appeared normal, whereas anthracis and Proteus mirabilis. It is potentially the drug 20 the left great toe was still moderately inflamed. This shows of choice with a secondary indication for use against improved germicidal activity of iodine with dimethyl infections by Bacteroides, Actinomyces, and Salmonella. sulfoxide. The use of DMSO with penicillin is illustrated in the EXAMPLE 42 following example: The fur was removed from a back area on four rabbits. EXAMPLE 38 Three iodine solutions were prepared which contained A 20-year old male with bilaterally-infected ingrown 2% iodine in 2.3% sodium iodide water, 46% aqueous toenails was treated with 90% dimethyl sulfoxide applied ethanol, and 95.7% dimethyl sulfoxide in water. Each topically to the right great toe and 90% dimethyl sulf test solution was applied to the exposed area of a different oxide plus 10 cc. of aqueous penicillin (i.e. penicillin 30 rabbit. Gross observation disclosed that skin discoloration G), containing 1 million units applied to the left great with the dimethyl sulfoxide was transitory relative to the toe. After two days, there was no inflammation or infection discoloration with the other solutions. The two water and in the toe treated with the dimethyl sulfoxide and penicil alcohol Solutions produced long-lasting surface stains. lin composition, whereas the toe treated with dimethyl Staphylococcus-infected sutures were implant by passing sulfoxide alone had a minimal subsidence of infection. 3 5 them through the entire skin and subcutaneous portion in The penicillin in the composition was therefore carried the treated area of each rabbit. The iodine sample treated across the skin barrier so that it contacted the infected with dimethyl sulfoxide composition applied topically infection site. These results show that dimethyl sulfoxide controlled the infection, whereas the aqueous solutions and enhanced penetration of the antibiotic. the ethanol solutions of iodine did not. The streptomycin group of antibiotics which includes 4) streptomycin, neomycin, kanamycin, paramomycin, and EXAMPLE 43 viomycin are bactericidal for a wide variety of bacteria, The four was removed from the anterior abdominal area including the tubercle bacillus, but must be used with on six rabbits. Two rabbits had 100% dimethyl sulfoxide care in view of their renal and central nervous system applied to the areas; two other rabbits had 2% tincture of toxicity. This group of drugs is effective against many iodine applied to the areas; and the other two rabbits had penicillin-resistant organisms. a composition applied which contained 2% iodine, 2.3% The tetracyclines, particularly tetracycline, oxytetra sodium iodide and 95.7% dimethyl sulfoxide. A full thick cycline chlortetracycline and demethylchlortetracycline, ness biopsy was taken from the exposed skin areas, the are the drugs of first choice in infections by Shigella, Samples transferred to a tube and cultured, and colony Brucella, Bacteroides, Eaton agent, and the Psittacosis 50 counts were taken. The colony counts ranged from 280 LGV-Trachoma viruses. They are possible drugs of choice 100 on the two rabbits which received the tincture of (secondarily indicated) in Klebsiella infections, hospital iodine alone. The counts were 20 to 30 on the animals borne coliform infections, and respiratory tract infections. receiving dimethyl sulfoxide alone and the counts were As previously indicated, DMSO may additionally act 2 and 3 respectively on the two animals receiving the com to increase the sensitivity of previously resistant strains position of iodine and dimethyl sulfoxide. of microorganisms to antibiotics. Following Examples 39 Illustrative of diseases caused by viruses are smallpox, and 40 illustrate this aspect: measles, encephalitis, herpes, rabies, etc. EXAMPLE 39 While many drugs are useful in treating viruses in vitro, A 36-year old female with an axillary abcess was very few are successful in treating in vivo virus disorders. attended surgically. The abcess was incised and cultured. 60 The most successful of the current antiviral agents are The findings were: Staphylococcus, coagulase positive, idoxuridine (5-iodo-2'-deoxyuridine), N-methylisatin beta resistant to penicillin, tetracycline and erythromycin, but thiosemicarbazone, and amantadine. Idoxuridine is useful sensitive to chloromycetin. An application of 50% aque in treating herpes simplex keratitis, and N-methylisatin ous dimethyl sulfoxide was made directly into the abcess beta thiosemicarbazone has proved useful in preventing three times daily. At the end of 48 hours and abcess was smallpox after exposure to this disease. recultured and showed staphylococcus, coagulase positive, EXAMPLE 44. sensitive to penicillin, tetracycline, erythromycin and chlo romycetin. It may be suggested that dimethyl sulfoxide The following lotion may be formulated incorporating alters sensitivity of the bacteria to antibiotics, perhaps by idoxuridene at a concentration 0.1 gm./cc. of the follow allowing an increased penetration of the antibiotic into the ing lotion base: bacterium. Percent EXAMPLE 40 Dimethyl sulfoxide ------75 Water ------14 A 60-year old male subject, with gangrene of the right Glycerine ------10 great toe, secondary to peripheral arteriosclerosis with Sodium carboxymethylcellulose ------3,551,554 25 26 The foregoing lotion may be employed in the treatment tion (the DMSO being employed either to enhance pene of herpes simplex by cutaneous application directly to tration of some other agent or for its own direct phar skin lesions, qi.d. macological effect) to suppress symptoms of histamine The primary disorders caused by protozoa are malaria release caused by the DMSO (e.g. urticaria, burning sen and amebiasis. The principal drugs employed in the treat sation, etc.). The following examples are illustrative. ment of malaria are quinine, quinacine, quinoline deriva 5 tives, and chlorophenyl derivatives. EXAMPLE 47 In the treatment of amebiasis, various drugs are em A twenty-two year old white male subject was treated ployed such as emetine, various arsenical amebacides, hy who had a three hour history of a common cold. He had droxyquinoline derivatives, antimalarial drugs and anti marked nasal congestion and discharge. Two mg. of di biotics, such as bacitracin, chlortetracycline hydrochlo O phenhydramine hydrochloride (Benadryl) in 0.5 cc. of ride, oxytetracycline hydrochloride, carbomycin, erythro normal saline was placed into each nostril. The symptoms mycin, paromomycin, and fumagillin. cleared, and the subject was asymptomatic for one and EXAMPLE 45 one-half hours. One half cc. of 50% dimethyl sulfoxide 15 in water was placed in each nostril. The symptoms cleared The following vaginal jelly may be prepared and buff and remained relieved for three hours. ered to a pH of about 4.0: After the symptoms of nasal congestion and discharge Percent returned, the subject was treated with 0.5 cc. of 50% di Ricinoleic acid ------0.8 methyl sulfoxide in an aqueous solution containing 2 mgs. Acetic acid ------0.75 20 of diphenhydramine hydrochloride. The symptoms were Boric acid ------3.0 quickly relieved and remained absent for twelve hours. Dimethyl sulfoxide ------15.0 EXAMPLE 48 Oxyquinoline Sulfate ------0.03 Sulfisoxazole (Gantrisin) ------8.0 A cyclizine hydrochloride mist formulation may be pre Tragacanth ------pared and charged into an aerosol container to provide Acacia ------fifty 100 mg. doses which may be administered intra Methyl paraben ------nasally for treatment of allergy states and motion sick Potassium hydroxide -- q.S. ness, etc. Five grams of cyclizine hydrochloride are in Potassium bitaritrate --- corporated in a halocarbon propellant formulation base Perfume ------. 30 containing 4% by volume DMSO. Water ------EXAMPLE 49 The foregoing vaginal jelly may be administered intra A topical ointment formulation of tripelennamine base vaginally as an average dose of 5 cc. in the evening before particularly suitable for treatment of itching dermatoses resting and again in the morning for the treatment of (applied to the affected area several times daily), may be Trichomonas vaginalis, formulated as follows: Various diseases are caused by yeasts. These include Percent moniliasis and North American blastomycosis. Antimicro Tripelennamine base ------2 bial agents employed in treating these diseases include DMSO ------70 nystatin, amphotericin B, sulfonamides and diamidines. 40 Sodium carboxymethylcellulose ------4 EXAMPLE 46 Water ------24 The following lotion for topical application to areas in Neuropharmacologic agents fected with yeast organisms may be prepared: "Neuropharmacologic agents' are those agents having a pharmacologic activity involving the nervous system. Dimethyl Sulfoxide ------IIl--. 80 45 Central nervous system active drugs have their site of CarboWax 1000 ------Ill-- 10 activity in the brain and/or spinal cord. Other agents have Water ------gll- 10 their activity in the peripheral nervous system. In every Nystatin (USP) ------units/cc. 100,000 case, the rapidity and magnitude of response to a neuro Antihistamines pharmacologic agent is dependent upon the speed and Antihistamines are agents which antagonize the pharma 50 extent of conduction or movement of the agent from the cological actions of histamine. They are employed in the site of administration to the site of activity in the nervous system. In many instances, the usefulness of such drugs is symptomatic treatment of various allergic states wherein limited by the inability of such agents to rapidly penetrate the antihistamine suppresses the symptoms attributable to membranes to achieve a useful concentration or distribu the actions of histamine released in the body in these dis 55 orders. Illustrative of such allery states are allergies of tion at the site of activity in the nervous system. The use the respiratory tract, for example, coryza, hay fever and of any neuropharmacologic drug must take into account vasomotor rhinitis, allergic dermatoses, angioedema and the problem of getting the drug from the periphery to the systemic allergies, for example serum sickness and drug action site which requires transport across neural barriers, reactions. Additionally, various chemicals and drugs cause 60 including the blood-brain barrier in the case of CNS histamine release in the body. agents. Antihistamines also have central nervous system activity Central nervous system active agents as sedatives and hypnotics. They are used in treating Central nervous system active agents are those agents nausea and vomiting (antiemetic use), insomnia and symp which stimulate, depress or otherwise modify central toms of parkinsonism. They also have local anaesthetic 65 nervous system function, either in a selective or non activity making them useful in control of itching. selective manner. Most of the central nervous system Antihistamines include the ethanolamines, e.g. diphen active agents described and illustrated under the following hydramine, the ethylenediamines, e.g. pyrilamine, the subheadings have useful activity in one or more supra alkylamines, e.g. chlorpheniramine, the piperazines, e.g. medullary portions of the brain, i.e., those portions lying chlorcyclizine, and the phenothiazines, e.g. promethazine. 70 above the medulla oblongata including the cerebrum, cere In addition, with regard to their use in treating allergy bellum, thalamus, and hypothalamus. An example is states, antihistamines may be used to counter the side aminophylline of Example 34 which is active in the cere effects due to histamine release caused by the application bral cortex. Agents which provoke a neuropharmacologic of DMSO to tissue. Notably, antihistamines may be in response in the central nervous system generally must pass corporated into DMSO formulations for topical applica 75 through the body from the site of administration, through 3,551,554 27 28 the blood stream, to the blood/brain barrier. To carry out or moving. This example illustrates potentiation of bar their activity, these agents then must pass through this biturate with dimethyl sulfoxide. barrier. Relatively little is known regarding passage of drugs into the brain from the blood stream, but in effect EXAMPLE 52. the “blood-brain barrier' behaves like a lipoid membrane. A chloral hydrate suppository formulation may be pre Most agents pass through this barrier with great difficulty. 5 pared by blending together: The effectiveness of a neuropharmacologic agent on the Gms. central nervous system is dependent upon the rate and Chloral hydrate ------10 extent of its penetration across this barrier. Therefore, in DMSO ------4.7 addition to enhancing penetration of such agents from the Stearic acid ------1. site of administration into the blood stream, dimethyl 10 Water ------0.5 sulfoxide may also play the role of enhancing penetration The melt is poured into a suppository mold and cooled of these agents across the blood-brain barrier. Following to form 10 suppositories each supplying a 1 gram dose. are illustrations of the various classes of central nervous One or two suppositories, as indicated, may be adminis system active agents with specific examples of the use of tered as a general sedative. dimethyl sulfoxide to enhance penetration of these agents: Analgesics and antipyretics CNS stimulants Agents in this category include strychnine picrotoxin, Analgesics exert type of depressing action upon the pentylenetetrazol and the xanthines (e.g. theophylline, central nervous system, the result of which is the obtunding theobromide and aminophylline). These agents generally of pain senations without the loss of consciousness. Some operate by the mechanism of blockage of inhibition (e.g. classes of analgesics, e.g. the salicylates, para-aminophenol strychnine) or by direct neuronal excitation (e.g. pentyl derivatives and the pyrazolon derivatives, also are anti enetetrazol). Preceding Example 34 and following Ex pyretics, that is, they lower abnormal body temperature amples 53 and 69 are illustrative: through a predominately CNS action. Additionally many 25 agents in these classes have other actions such as anti EXAMPLE 53 inflammatory action. The para-aminophenol derivatives include acetanilid, acetophenetidin and acetaminophen. A unit dose suppository form of imipramine can be Exemplary of the salicylates are acetylsalicylic acid and prepared by melting together: salicylic acid and their non-toxic salts, esters and related 30 Imipramine base ------mg - 50 derivatives. The pyrazolon derivatives include antipyrine DMSO ------gm-- 4.5 aminopyrine and phenylbutazone. The narcotic analgesics Sodium stearate ------gm-- 1.0 (opiates) include morphine, codeine, meperidine, opium, Glycerine ------gm-- 4.5 etc. Preceding Example 34, and following Examples 50 Water ------1.0 and 67 are illustrative: and cooling the melt in a suppository mold. This dosage EXAMPLE 50 may be administered rectally T.I.D. to relieve depression. The following injectible formulation is prepared in 2 cc. Anticonvulsants and centrally acting muscle relaxants ampules to provide doses of 1-2 cc. for intramuscular in The various CNS-active agents used to treat convulsive jection (q.i.d. for sustained relief): 40 disorders, particularly the various types of epilepsy, in Meperidene hydrochloride ------mg-- 250 clude the barbiturates, glutarimides, hydantoins, acetyl DMSO ------gm-- 1.5 ureas, oxazolidinediones and the succinimides. Cen Water ------gm-- 8.5 trally acting muscle relaxants have the ability to diminish Sodium acetate ------gm-- 0.25 skeletal muscle tone and involuntary movement by action Sodium formaldehyde sulfoxylate ------gm-- 1 on the CNS. They are used as an adjuvant in general General CNS depressants anaesthesia and in treatment of alcoholism, parkinsonism and cerebral palsy. General CNS depressants act nonselectively to depress Such agents include mephenesin, methocarbamol, sty all excitable tissue, in general, through stabilization of ramate, chlorZoxazone, carisoprodol, metaxalone, tri part or all of the neuronal membrane. They are utilized hexyphenidyl and benztropine mesylate. The following as general anaesthetics, hypnotics and sedatives. Promi example is illustrative: nent among these agents are the barbiturates and related sedative-hypnotics such as the chloral derivatives, bro EXAMPLE 54 mides, tertiary acetylenic alcohols, carbamic acid esters The following parenteral formulation may be prepared of alcohols and glycols (e.g. ethinamate and meproba 5. 5 and sterilized for treatment of grand mal and psycho mate), monoureides, diureides, benzodiazepines, piperi motor epilepsy to achieve more rapid therapeutic benefit. dinedione derivatives, etc. The following examples are illustrative: Diphenylhydantoin ------mg-- 250 DMSO ------ml.-- 4 EXAMPLE 51 60 Water ------ml -- 1 Three groups of dogs were evaluated. Three dogs in The dose is 1-3 cc. preferably administered intramus group No. 1, weighing about 15 to 18 kgs., were given cularly. 1 cc. containing 50 mgs. of sodium pentobarital (Nem butal) per 5 pounds of body weight, intravenously. This Psychopharacological drugs produced a profound anaesthetic effect, allowing an ab Agents used to treat psychiatric disorders fall into sev dominal operation without causing the dog any discom eral categories. Drugs used to treat anxiety and neurotic fort. Another group of three dogs were given 1 cc. of conditions include the benzodiazepine derivatives, mepro Nembutal per 10 pounds of body weight. These dogs re bamate and various other non-barbituate and barbituate mained partially awake and surgery could not be per sedatives which have been treated previously concerning formed without causing discomfort to the dogs. The last their employment in the present invention, under the group of three dogs were given 1 cc. of Nembutal per 10 heading of general CNS depressants. The predominant pounds of body weight plus 1 gram of dimethyl sulfoxide drugs employed to treat psychoses are primarily the per 2 pounds of body weight, instilled orally through a phenothiazines and the rauwolfia alkaloids. In treatment gastric tube. After one-half hour, surgery could be per of depression, monoamine oxidase inhibitors and dibenz formed without the animal experiencing any discomfort - azepine derivatives are the predominant drugs. Pre 3,551,554 29 30 ceding Example 53 and the following example illustrate EXAMPLE 58 the use of DMSO to enhance penetration of these agents: The following solutions were prepared: EXAMPLE 55 Solution A The following formulation may be melted together, Cc. placed in a suppository mold and cooled to form a unit Lidocaine hydrochloride (2%) ------2 dose for rectal application as a monoamine oxidase in Trisodium phosphate ------2 hibitor for treatment of depressive states: Glycerine ------: earner - - en man 2 N-benzyl-N-methyl-2-propynylamine ------mg-- 100 Solution B DMSO ------gm-- 4 O Cc. Diglycol laurate ------gm-- 1.5 Lidocaine hydrochloride (2%) ------2 Carbopol 934 ------gm-- 0.25 Trisodium phosphate buffer ------2 Triethanolamine ------gm-- 0.01 Dimethyl sulfoxide ------2 Peripheral nervous system active agents Solution A was administered topically to the general Agents which have activity in the peripheral nervous area overlying the mental foramen of one human subject, system include local anaesthetics which reversibly block and solution B was also administered topically to the nerve conduction when applied locally to nerve tissue, mental foramen of another human subject. The general thereby to relieve pain. They also include agents which area overlying the mental foramen is in midline of the have activity at various sites in the autonomic nervous 20 exterior surface of the mandible. system, including anticholinesterase agents, parasym Solution B produces sufficient local anaesethesia within pathomimetic agents, sympathomimetic agents, antimus fifteen minutes to permit a lower bicuspid tooth with carinic agents, adrenergic blocking agents, ganglionic carious lesions to be drilled without discomfort. The blocking and stimulating agents and neuromuscular block subject receiving solution A did not develop such a level ing agents. Following is a discussion and illustration of 25 of anaesthesia within the same time limit. the employment of DMSO with these classes of agents EXAMPLE 59 in accordance with the invention. A forty year old female subject with pruritis ani was Many compounds which produce local anaesthesia have given a topical application of 3 cc. of 100% dimethyl a common fundamental structure, namely, a hydrophilic sulfoxide to the irritated area. Relief of itching occurred amino group (a tertiary or secondary amine), connected 30 in fifteen minutes and lasted for eight hours. The sub by an intermediate group through an amide bond oester ject complained of a mild "burning sensation” in the linkage to a lipiphilic aromatic residue. Representative area of application which lasted for ten minutes. The of these agents are procaine, lidocaine, piperocaine, di same burning sensation was reported after each of four bucaine, tetracaine, cyclomethycaine, pramoxine and di applications over a two-day period. methisoquin. Others include benzocaine, orthoform, butyl 35 A 4% xylocaine solution in 100% of DMSO was made. aminobenzoate, benzyl alcohol, methol, phenol and qui Four cc. of the mixture were applied to the same sub nine. Antihistamines have also been employed for this ject who reported similar relief of discomfort with no purpose. Because of their low solubility in water and "burning.” This example shows that small amounts of other systems in their active form, local anaesthetics are 40 a local anaesthetic will lessen the discomfort which may frequently utilized in the form of water soluble salts, occur with dermal application of high concentrations of rather than as free bases. Advantageously DMSO may dimethylsulfoxide. permit use of the free base form when its is employed EXAMPLE 60 as a solvent for the local anaesthetic in addition to being utilized as a penetrating agent. Employment of local Two solutions were prepared, one containing 0.44% anaesthetics with DMSO may provide a special advantage Xylocaine base in 100% dimethyl sulfoxide and the other in relieving the burning sensation frequently experienced containing 0.5% menthol in 100% dimethyl sulfoxide. when DMSO is applied topically for its own pharmaceuti Human subjects who previously reported a "burning sen cal effects or for enhancing the penetration of yet another sation' with 100% dimethyl sulfoxide reported absence pharmaceutical agent. The following examples are il of such a "burn' when they were given like amounts of lustrative: 50 the two solutions in a like dermal application. EXAMPLE 56 EXAMPLE 61 A 1% procaine hydrochloride isotonic solution was Neostigmine, one of the most prominent anticholine administered by subcutaneous method in 0.5 mm. doses sterase agents, may be formulated with DMSO in single in the human skin. In another human subject 1% procaine 55 dosage form as follows: in isotonic solution containing 15% v./v. of dimethyl Neostigmine base ------Ing-- 0.5 sulfoxide was administered by subcutaneous injection in DMSO ------ml.-- 0.7 a 0.5 mm. dose. The dimethyl sulfoxide composition pro Water ------ml.-- 0.3 duced a local anaesthetic effect having a prolonged activ Methyl propyl paraben ------gm-- 0.0002 ity of 1.5 times procaine alone. 60 This formulation may be packaged in 1 cc. ampules EXAMPLE 57 each of which is then diluted with 4 cc. of distilled water Isolated peripheral cutaneous nerves of dogs were stim for bladder instillation as a preventative of post-operative ulated with electrodes to measure impulse conduction. Ap distention and urine retention. The dosage is repeated plication of 25% aqueous dimethyl sulfoxide to a first 65 every 4-6 hours as needed. nerve provided a local and reversible block of impulse EXAMPLE 62 conduction which lasted twenty minutes. Application of To demonstrate enhanced penetration of the sympath a 2% procaine hydrochloride solution blocked impulse omimetic agent epinephrine, epinephrine hydrochloride conduction for forty-five minutes in a second nerve. Ap 70 is dissolved in 90% aqueous DMSO to a concentration plication of an aqueous solution containing 25% dimethyl of 0.05% and applied topically to the stretched ear of sulfoxide and 1% procaine to a third nerve blocked im a hamster fixed in position for low power microscope pulse conduction for one and one-half hours. This shows viewing of the capillary bed. Whereas epinephrine hydro and enhanced effect when procaine is combined with di chloride dissolved in water (at the same concentration) methylsulfoxide. 75 applied to the hamster ear produces no visible change 3,551,554 31 32 in appearance of the capillary bed, with the DMSO form EXAMPLE 67 ulation pronounced constricture of the capillary bed is observed by microscope and a general blanching of skin The following phenylbutazone topical formulation may is seen. This effect is transistory. The same DMSO form be prepared and applied in unit doses of 10 cc. bid topi ulation may be used as a nasal spray for relief of asth cally to the involved area to treat musculoskellatan pain matic attacks. and inflammation: Gms. EXAMPLE 63 Phenylbutazone ------6 The sympathomimetic agent, methamphetamine may DMSO ------90 be formulated in a unit dose with DMSO for intramuscu O HaO ------10 lar injection as follows: Anticoagulants Methamphetamine ------19- 20 DMSO ------ml.-- 0.3 Anticoagulants are agents used primarily in thrombo USP Water ------ml.-- 0.7 embolic conditions to prevent formation of intra-vascular Propyl paraben ------gm - 0.0001 thrombi and to maintain normal hemostasis. Heparin and its soluble metal salts, the most quick acting anticoagul The product is packaged in a 1 cc. ampule for admin lants, previously required parenteral administration so istration to maintain blood pressure in hypotensive states the employment of DMSO to permit topical application as during spinal anaesthesia. is of considerable advantage. The coumarin and indand 20 ione derivatives, e.g. bishydroxycoumarin and henindione, EXAMPLE 64 may be administered orally. However, in some circum Mecamylamine is representative of agents having a stances, their topical application may be advantageous. site of activity at the autonomic ganglia (ganglionic stim When applied topically with DMSO, these agents may be ulating and blocking agents). It may be formulated as penetrated into the general circulation to provide the de follows to provide a single dose in suppository form for sired blood level. Reference is made to foregoing Exam rectal application in treatment of hypertension: ple 34 and to the following example for illustration of the use of DMSO to enhance penetration of this class of Mecamylamine base ------mg-- 15 agents: Propylene glycol Stearate ------gm-- 4.5 30 DMSO ------gm-- 4.5 EXAMPLE 68 Triethanol amine ------gm.-- 0.3 Stearic acid ------gm-- 1.5 Forty thousand units of heparin (1 cc.) with 1 cc. 100% dimethyl sulfoxide was placed onto human skin Melt ingredients together and pour into cooled sup (left forearm-thirty nine year old male). A Lee and pository mold. White 3 tube clotting time was taken prior to application. EXAMPLE 65 This was 11 minutes. Clotting times were taken at 2 hours and 4 hours subse Also exemplary of adrenergic agents is the adrenegic quent to administration. At 2 and 4 hours, the clotting blocking agent dihydroergotamine. A topical lotion may 40 times were 14 and 18 minutes respectively. This example beformulated as follows: shows that dimethyl sulfoxide was associated with ab Dihydroergotamine ------mg. 200 sorption of heparin through human skin. DMSO ------ml. 180 Water ------ml.-- 18 Vasodilators Disodium hydrogen phosphate ------gm.-- 2 Vasodilators are generally used clinically in treatment Two cc. of this preparation may be applied topically of ischemic conditions, especially myocardial hypoxia, in treatment of herpes zoster for relief of neuritic pain as in angina pectoris, and to some extent in ischemia or for peripheral vasodilation in arterial deficiencies. of skeletal muscle. Their basic pharmacological action is the relaxation of smooth muscle to effect dilation. These EXAMPLE 66 agents include the nitrides, such as sodium nitrite, glyceryl tri-nitrate and isosorbide di-nitrate, di-pyrida Representative of parasympathomimetic agents is pilo mole, cyclandelate, nicotinic acid and aminophylline. carpine. The following eye drop formulation may be re DMSO may be utilized to promote penetration for topical pared: application of these agents to provide the desired sys Gms. 5 5 temic concentration. The following examples and fore Pilocarpine HCl ------3 DMSO ------O going example 34 illustrates the use of DMSO to en Isotonic Saline ------86.5 hance penetration of this class of agents: Sodium dihydrogen phosphate ------0.5 EXAMPLE 69 to which is added 0.01% benzalkonium chloride as a 60 preservative. Aminophylline is a known diuretic, central nervous sys tem stimulant, coronary vasodilator and bronchodilator A typical dose for the treatment of chronic open-angle which is normally absorbed across the bladder walls. glaucoma is 1-2 drops in the eye every 4-6 hours. A test solution of phosphate buffer containing 0.033 M. Anti-inflammatory agents 65 phosphoric acid/100 ml. and 0.05 M. trisodium phos phate/100 ml, containing 15% dimethyl sulfoxide, was Anti-inflammatory agents are agents which diminish the compared with a control solution of the same buffer solu inflammatory response due to tissue injury. They com tion. The pH of both buffer solutions was 9. The control prehend the corticosteroids considered previously as well and test solutions were instilled in the freshly evacuated as most of the analgesics and antipyretics discussed un 70 bladders of two dogs in an amount of 75 cc. Blood sam der that heading (e.g. the salicylates and pyrazolon ples were taken at different time periods for each dog, derivatives). For illustration of use of DMSO to en and analysis for aminophylline by the Brakett and Brad hance penetration of these agents reference is made to ford test was done. The results in the following table preceding Examples 2, 3, 4 and 34 and the following ex showed an increased amount of aminophylline in the ample: blood of the dog receiving the test solution, 3,551,554 33 34 AMINOPEIYLLINE SERUMLEVELNDOGS (MG. PERCENT) sulfonate is injected for localizing ureteral orifaces. Average Cationic dyes are used for in vitro tissue staining to fix conce cellular structure and the like for microscopic observa Average tration tion and evaluation. Radiopaque media are used in concen- 15% di tration methyl roentgenography of various body structures, usually ad control sulfoxide ministered by injection. Preceding Example 34 and the buffer in buffer following examples illustrate the use of DMSO to en Time in minutes post injection: hance tissue penetration of diagnostic dyes and radiopaque 30. 1.7 2.8 60- 2.17 4.6 media: 120. 4.03 5.9 EXAMPLE 73 240. 3.77 5.75 O A solution of 2% basic green (MX) dye in 100% di A group of two rats received 100 mgs. of nicotinic methyl sulfoxide was applied to the freshly washed skin acid in 4 cc. of 100% dimethyl sulfoxide applied to the of the forearm. The same dye in a like concentration in anterior abdomen. A wet state of application was main Water, ethanol, acetone and ethylene glycol, was also tained. A control group of two rats received 100 mgs. of 5 applied to freshly washed areas of the forearm. The excess nicotinic acid in 4 cc. of saline. At the end of two hours, dye was removed with detergent or water, and the extent the test group showed noticeable vasodilation of the skin and depth of the dyeing was determined by stripping away with an increase in cutaneous temperature. These obser approximately 0.5 mm. thick layer of skin with adhesive vations were absent in the control group. This example tape. The dye and dimethyl sulfoxide was found to be the shows increased penetration with dimethyl sulfoxide of 20 only composition which carried the dye to the deeper an agent which is both a vasodilator and a vitamin. dermal layers. This shows the enhanced dye absorption when administered in combination with dimethyl sulf EXAMPLE 71 oxide. A glycerol trinitrate ointment may be prepared by EXAMPLE 74 blending the following ingredients: 25 Gms. A 25% solution of iodine in aqueous sodium iodide was Glycerol trinitrate ------2 prepared as a first diagnostic solution, and a 25% solution DMSO ------70 of iodine in 100% dimethyl sulfoxide was prepared as a Ethanol ------8 second diagnostic solution. Twenty ml. of each solution CarboWax 1500 ------20 30 was injected into the intermedulla of femur bones. The A typical dosage of 10 mg. of glycerol trinitrate is first solution was injected into the right femur, and the provided with /2 gram of this ointment. The dosage second into the left femur of the same subject. X-ray may be applied topically as, for example, to the intact photographs were taken at 5-minute sequences over 240 skin of the upper arm, or Sublingually. minutes of both femurs. After 40 minutes, a picture of the left femur, into which was injected the second solution, Ultraviolet Screening agents showed the entire intermedullar space to be radiopaque. Compounds which screen a part or all ultraviolet light At the same time, the first solution in the right femur was from the skin to protect the skin from the damaging rapidly diffusing into the vascular system and being re effects of over-exposure to sunlight are called ultraviolet moved from the bone. At 120 minutes, a picture of the left absorbers or screening agents. They are applied topically 40 femur was still clearly defined as being radiopaque. At the to the skin prior to exposure to absorb the ultraviolet rays same time, a picture of the right femur showed no evi to thereby prevent injury to the skin. Examples of such dence of opacity except for about a one centimeter circle agents are 2-ethoxyethyl p-methoxy cinnamate, digalloyl around the injection site. trioleate, menthyl anthranilate, p-aminobenzoic acid, Nutrients phenyl salicylate, and the benzophenones, e.g. 2,4-dihy 45 droxybenzophenone and 2,2'-dihydroxy-4,4'-dimethoxy As used herein the term nutrients is intended to com benzophenone. DMSO may be utilized to enhance penetra prehend the vitamins, carbohydrates, fats, proteins, includ tion of these agents into the epidermis, thereby to increase ing proteinaceous hormones and mineral nutrients used their effectiveness and to increase their lasting power by by the body to sustain and regulate metabolism and inhibiting their accidental removal from the skin through 50 furnish energy. Exemplary of the carbohydrates are bathing, perspiration, etc. The following example is illus glucose ( see Example 1) and dextrins, (e.g. maltose trative: dextrins). Metabolizable fats are the glyceryl esters of EXAMPLE 72 fatty acids, e.g. vegetable oils. The various commercial A male subject had a skin markedly sensitive to ultra protein hydrolyzates, e.g. hydrolyzed casein, and insulin violet light. A test solution was made containing 1% ultra 55 are representative of the proteins. The vitamins include violet absorber in 100% dimethyl sulfoxide. A control the water-soluble vitamins, e.g. vitamins B1, B2, B6, and solution was made containing the ultraviolet absorber in B12, nicotinic acid, pantothenic acid and p-aminobenzoic 100% ethanol, and both solutions were similarly thickened acid and the fat soluble vitamins, e.g. vitamins A, D, K. with Carbowax 4000 to provide lotion forms. The ultra and E and folic acid. Mineral nutrients include inorganic violet absorber was 2,2'-dihydroxy-4,4'-dimethoxy benzo 60 salts of fluorine, iodine, manganese, potassium, iron, zinc, phenone. The two lotions were applied to different sides copper magnesium calcium and phosphorous. DMSO is of the subject's face. After one day of severe sun ex useful in enhancing tissue penetration of nutrients for as posure, the subject was examined. The side with the con simulation by the body particularly by the topical or in trol application showed marked redness. The side with jection routes. Preceding Examples 1 and 70 and the fol the test application showed only slight redness. Two days 65 lowing additional examples are illustrative: later, the control side was blistered, whereas the test EXAMPLE 75 side was normal and free of redness. Two dogs of approximately 15 kgs. of body weight were Diagnostic dyes and radiopaque media starved for twenty-four hours and administered three units Tissue compatible cationic dyes are employed in tissue 70 of crystalline zinc insulin subcataneously. Another group staining both in vivo and in vitro for diagnostic proce of two dogs was starved for twenty-four hours and ad dures. For use in vivo, this may be for marking or follow ministered three units of insulin subcutaneously in 1 cc. ing visually the tissue penetration of another agent or of 100% dimethyl sulfoxide. The dogs given the composi for highlighting or outlining a particular organ or struc tion including the dimethyl sulfoxide were observed to ture for visual diagnosis. For example, sodium indigotindi 75 show signs of severe insulin shock two hours after ad 3,551,554 35 36 ministration. The insulin without dimethyl sulfoxide did agents and nutrients, which comprise the concurrent topi not produce a pronounced physiological response. cal administration to the external membrane of an amount of said agent effective to produce the desired physiological EXAMPLE 76 effect and an amount of DMSO Sufficient to effectively Four dogs with pancreatectomy were given insulin enhance penetration of said agent to achieve the desired orally. A group of two dogs were used for controls and 5 physiological effect. were given 15 units of insulin in 3 cc. of isotonic saline, 2. A method as in claim 1 and wherein the said agent and the other group of two dogs was given 15 units of is applied to the intact skin in a composition which in insulin in 3 cc. of 99% dimethyl sulfoxide. Blood sugars cludes said DMSO and wherein the DMSO in said com Were taken at one, two and four hours. The two dogs in O position is at least about 50% by weight of the composi the control group started with blood Sugars of 82 mg. tion. percent and 90 mg. percent. At four hours, the blood 3. A method as in claim 2 and wherein said agent is Sugars had risen to 100 and 110 mg. percent. an antihistaminic agent. The two animals in the test group receiving the di 4. A method as in claim 1 and wherein said agent is methyl sulfoxide started with blood sugars of 80 mg. applied to mucous membrane of a body cavity in a com percent and 85 mg. percent, and at the end of two hours, position which includes said DMSO and wherein the one had 60 mg. percent and the other 65 mg. percent. DMSO in said composition is at least about 10% by At the end of the four hours, one had 40 mg. percent weight of the composition. and the other 35 mg. percent. It is believed that the com 5. A method as in claim 1 and whereby said agent is position containing dimethyl sulfoxide enhanced pene selected from the group of compounds consisting of cat tration of the insulin through the esophageal wall, since ionic compounds, anionic compounds and non-dissociat insulin is known to be destroyed in the stomach. ing compounds. 6. A method as in claim 5 and wherein the molecular EXAMPLE 77 weight of said agent is less than about 8000. A group of two rats received 0.5 gms. of ammonium 7. A method as in claim 1 and wherein said agent is fluoride in 4 cc. of 100% dimethyl sulfoxide by applying an antineoplastic agent selected from the group consisting this composition over the abdomen. Another group of of alkylating agents and antimetabolites. two rats Served as controls, receiving 0.5 gms. of am 8. A method as in claim 1 and wherein said agent is monium fluoride in 4 cc. of isotonic saline. A wet system applied to said membrane in a composition which includes was maintained. The test group receiving dimethyl sul 30 said DMSO. foxide and the ammonium fluoride showed typical epi 9. A method as in claim 8 and wherein said agent is leptiform convulsions associated with the absorption of a neuropharmacological agent having useful activity in large amounts of fluoride. The control group did not show the central nervous system. Such convulsions. This example shows that dimethyl sul 10. A method as in claim 9 and wherein said agent is an foxide enhanced absorption of ammonium fluoride analgesic. through intact skin and therefore may increase fluoride 11. A method as in claim 8 and wherein said agent is a absorption into teeth. neuropharmacological agent having activity in the periph" eral nervous system. EXAMPLE 78 12. A method as in claim 11 and wherein said agent An ointment base may be prepared from the following: 40 is a local anaesthetic. Parts by wt. 13. A method as in claim 8 and wherein said com Lanolin ------90 position contains a pharmaceutically acceptable thicken DMSO ------O ing agent in an amount sufficient to materially increase Isopropyl myistate ------5 the viscosity thereof, whereby to facilitate topical appli cation. to which is added 1800 U.S.P. units of Vitamin A and 14. A method as in claim 13 wherein Said composition 300 U.S.P. units of Vitamin D per gram of ointment is in the form of an ointment. base. This ointment is applied topically for the treatment 15. A method as in claim 13 and wherein Said Com of burns, skin irritation, diaper rash and pruritis. position is in the form of a lotion. EXAMPLE 79 16. A method as in claim 13 and wherein Said Com Tests were done with monkey kidney cell tissue cul position is in the form of a suppository. tures containing conventional tissue culture media (con 17. A method of enhacing the tissue penetration of taining glucose, minerals and amino acids). Two groups an injectable chemical agent capable of eliciting a physi of tests were made with replicates. To one group of ological effect in a human or animal subject, said agent tissue culture tests was added 0.003% dimethyl sulfoxide, 5 5 being seleted from the group consisting of antineoplastic and the other group was retained as a control. Cell counts agents, antihistaminic agents, neuropharmacologic agents were performed after three days, and control groups aver having a useful activity in a Supramedullary portion of aged 1,683,000 per cc., whereas the test group with di the brain, central nervous system depressants, analgesics, methyl sulfoxide tubes averaged 6,804,000 cells per cc. local anaesthetics, antiinflammatory agents, anticoagul Enhanced cell proliferation, therefore, occurred in cul 60 lants, vasodilators, diagnostic dyes, diagnostic radiopaque tures containing dimethyl sulfoxide. The growth is be agents and nutrients, which comprises the concurrent lieved to have occurred from greater utilization of the administration to said subject of an injected amount of nutrient in the tissue cultures containing dimethyl sulf said agent effective to produce the desired physiological oxide. effect and an amount of DMSO effective topically or by What I claim is: injection to enhance penetration of said agent into Said 1. A method of enhancing the penetration into and tissue to achieve the desired physiological effect. across an external membrane barrier of a human or 18. A method as in claim 17 and wherein said DMSO animal subject of a chemical agent capable of eliciting is administered by injection in a composition containing a physiological effect upon topical application thereof, at least about 1% by weight of DMST. said agent being selected from the group consisting of 19. A method as in claim 17 and wherein said agent non-estrogenic, antineoplastic agents, antigens, antihis is administered by injection in a composition containing tamic agents, neuropharmacologic agents, antiinflamma between about 1% and 40% by weight of DMSO. tory agents, anticoagulants, vasodilators, ultra-violet 20. A method as in claim 17 and wherein Said agent screening agents, diagnostic dyes, diagnostic radiopaque 5 is selected from the group of compounds consisting of 3,551,554 37 33 cationic compounds, anionic compounds and non-dissoci and said DMSO are injected together in the same com ating compounds. position. 21. A method as in claim 17 and wherein the molecu 40. A method as in claim 39 and wherein said agent lar weight of said agent is less than about 40,000. is a vasodilator. 22. A method of enhancing the penetration of a cati 41. A method as in claim 39 and wherein said agent onic dye through cell membranes of animal cells for di 5 is an anticoagulant. agnostic purposes which comprise applying to said cell 42. A method as in claim 39 and wherein said agent membranes a composition comprising an amount of said is an antiinflammatory agent. dye effective to stain said cells and an amount of DMSO effective to enhance penetration of said dye into said cells. 10 References Cited 23. A method of enhancing the penetration of a cat UNITED STATES PATENTS ionic dye through cell membranes of animal cells for diagnostic purposes which comprise applying to said cell 2,942,008 6/1960 Lubowe ------. 252-364 membranes a composition comprising an amount of said 3,044,936 7/1962 Achelis et al.------424-3373x dye effective to stain said cells and an amount of DMSO 5 3,067,096 12/1962 Trace et al. ------424-337)x effective to enhance penetration of said dye into said cells. FOREIGN PATENTS 24. A method as in claim 8 and wherein said agent is an antiinflammatory agent. 810,377 3/1959 Great Britain. 25. A method as in claim 8 and wherein said agent 234,383 9/1959 Australia. is a vasodilator. 20 OTHER REFERENCES 26. A method as in claim 8 and wherein said agent Ashwood-Smith: Int. Jl. Rad. Biol. 3(1): 41-48 Jan is an allergen. uary 1961, “The Radioprotective Action of Dimethyl 27. A method as in claim 8 and wherein said agent Sulfoxide and Various Other Sulfoxides.' is an infective antigen. Uranuma: Igaku Kenkyu 30: 2235-2261 (1961) "Tox 28. A method as in claim 8 and where in said agent icity of Dimethylsulfoxide as a Solvent.” is an antihistamine. Faust: American Perfumer 77 (1): 23-26 January 29. A method as in claim 8 and wherein said agent 1962, "Some New Components for Cosmetic and Derma is a nutrient. tological Vehicles.” 30. A method as in claim 29 and wherein said nutrient Marson: Boll. Chimico Farm. 102: 109-124 February is a vitamin. 30 1963, "Dimethyl Sulfoxide: A Waterminetic Solvent.’ 31. A method as in claim 30 and wherein said vitamin Rosenkranz et al: Cancer Chemotherapy Reports 31: is a water soluble vitamin. 7-24 September 1963, "Dimethyl Sulfoxide: Its Steroid 32. A method as in claim 9 and wherein said agent Solubility and Endocrinologic and Pharmacologic-Toxi is a central nervious system depressant. cologic Characteristics.” 33. A method as in claim 9 and wherein said agent Brown et al.: J. Pharm. Pharmacol. 15 (10): 688-692 . useful activity in a supramedullary portion of the October 1963, "A Note on the Toxicity and Solvent Prop 1. erties of Dimethyl Sulfoxide.” 34. A method as in claim 17 and wherein said agent Federal Register: 30 (228): 14639 Nov. 25, 1965, “Di is a neuropharmacologic agent having a useful activity 40 methyl Sulfoxide (DMSO) Preparations.” in a Supramedullary portion of the brain. Federal Register 33 (176), p. 12776, Sept. 10, 1968, 35. A method as in claim 17 and wherein said agent is "Dimethyl Sulfoxide (DMSO) Preparations.” a central nervous system depressant. 36. A method as in claim 17 and wherein said agent SHEP K. ROSE, Primary Examiner is an analgesic. 45 37. A method as in claim 17 and wherein said agent U.S. Cl.X.R. is an antineoplastic agent. 195-1.8; 424-4, 7, 9, 12, 45, 59, 88, 89, 91, 92, 128, 38. A method as in claim 37 and wherein said antineo 141, 144, 147, 150, 151, 153, 154, 170, 180, 181, 183, plastic agent is selected from the group consisting of 195, 200, 201, 209, 227, 228, 230, 236, 237, 238, 239, albylating agents an antimetabolites. 50 240, 242, 243, 244, 249, 250, 251, 253, 254, 255, 261, 262, 263, 266, 267, 269, 271, 273, 274 284, 285, 310, 39. A method as in claim 18 and wherein the agent 319, 320, 321, 322, 324, 330, 337, 344.