OA11307A - Drug delivery devices and methods for treatment ofviral and microbial infections and wasting syndro mes - Google Patents

Abstract

Novel drug delivery devices, methods and therapeutic compositions are described for treating viral and microbial infections and wasting syndromes in an animal, including a human patient. According to the invention, a polar compound such as dimethylformamide or dimethylsulfoxide is administered to a patient in need of treatment, preferably by a transdermal route. The invention further provides a vaccine prepared from antibodies harvested from the body of a patient treated by the method of the invention for a viral infection.

Description

011307

DRUG DELIVERY DEVICES AND METHODS FOR TREATMENT OF VIRALAND MICROBIAL INFECTIONS AND WASTING SYNDROMES

1. FIELD OF THE INVENTION

The présent invention is directed to methods of treating an animal affected by a viral or microbial infection, particularly an infection with a retrovirus, such as HIV, or by a wasting syndrome, especially a wasting syndrome associated with HIV infection or malignancy, by administering a polar compound such as N,N'-? dimethylformamide (DMF) or dimethylsulfoxide ÇDMSO) . Theinvention also provides pharmaceutical préparations anddrug delivery devices comprising a polar compound such asDMF or DMSO for treatment of an animal affected by a viralor zOther microbial infection or a wasting syndrome.

2. BACKGROUND OF THE INVENTION

2.1 THE HUMAN IMMUNODEFICIENCY VIRUS

Human immunodeficiency virus (HIV) induces. apersistent and progressive infection leading, in the vastmajority of cases, to the development of the acquiredimmunodeficiency syndrome (AIDS) (Barre-Sinoussi et al.,1983, Science 220: 868-870; Galla et al., 1984, Science224:500-503). There are at least two distinct types ofHIV: Hiv-l (Barre-Sinoussi et al., 1983, Science 220:868-870; Gallo et al., 1984, Science 224:500-503) and HïV-2(Clavel et al., 1986, Science 223:343-346; Guyader et -al., 1987, Nature 326:662-669). In humans, HIV réplication occurs prominently in CD4 + T lymphocyte populations, and HIV infection leads to déplétion of this cell type and eventually to immune incompétence, opportunistic 011307 infections, neurological dysfunctions, neoplastic growth,and ultimately death. HIV is a member of the lentivirus family ofretroviruses (Teich et al., 1984, RNA Tumor Viruses, Weisset al., eds., CSH-press, pp. 949-956). Retroviruses aresraall enveloped viruses that contain a single-stranded RNAgenome, and replicate via a DNA interraediate produced by avirally-encoded reverse transcriptase, an RNA-dependent DNApolymerase (Varmus, H., 1988, Science 240:1427-1439).

Other retroviruses include, for example, oncogenic virusessuch as human T-cell leukemia viruses (HTLV-1, -II, -III),and feline leukemia virus.

The HIV viral particle.. consists of a viral core,composed in part of capsid proteins designated p24 and pl8,together .with the viral RNA genome and those enzymesrequired for early réplicative events. Myristylated gagprotein forms an outer viral shell around the viral core,which is, in turn, surrounded by a lipid membrane envelôpederived from the infected cell membrane. The HIV envelôpesurface glycoproteins are synthesized as a single 160kilodalton precursor protein which is cleaved by a cellularprotease during virai budding into two glycoproteins, gp41and gpl20. gp41 is a transmembrane glycoprotein and gpl20 is an extracellular glycoprotein which remains non-covalently associated with gp41, possibly in a trimeric ormultimeric form (Hammerskjold, M. and Rekosh, D., 1989,

Biochem. Biophys. Acta 989:259-280). HIV, like other enveloped viruses, introduces viralgenetic material into the host cell through a viral-envelope mediated fusion of viral and target membranes.HIV is t-argeted to CD4+ cells because a CD4 cell surfaceprotein (CD4) acts as the cellular receptor for -the HIV-1virus (Dalgleish et al., 1984, Nature 312:763-767;Klatzmann et al., 1984, Nature 312:767-758, Maddon et al.,1985, Cell 47:333-348). Viral entry into cells is' 011307 3 dépendent upon gpl20 binding the cellular CD4 receptormolécules (Pal et al., 1993, Virology 194:833-837,- McDougalet al., 198S, Science 231:382-385, Maddon et al·., 1986,

Cell 47:333-348), explaining HIV's tropism for CD4- cells, 5 while gp41 anchors the envelope glycoprotein complex in theviral membrane. The binding of gpl20 to CD4 inducesconformational changes in.the viral glycoproteins, but thisbinding alone is insufficient to lead to infection(reviewed by Sattentau and-Moore, 1993, Philos. Trans. R. 10 Soc. London (Biol.) 342:59-66-).

Studies of HIV-1 isolâtes hâve revealed a heterogeneity in their ability to infect different humancell types (reviewed by Miedema et al., 1994, Immunol. Rev.140:35-72). The majority of extensively passaged 15 laboratory strains of HIV-1 readily infect cultured T cellsUnes and primary T lymphocytes, but not primary monocytesor macrophages. These strains are termed T-tropic. T-tropic HIV-1 strains are more likely to be found in HIV-1infected individuals during the late stages of aids (Weiss 20 et al., 1996, Science 272:1885-1886). The majority ofprimary HIV-1 isolâtes (i.e., viruses not extensivelypassaged in culture) replicate efficiently in primarylymphocytes, monocytes and macrophages, but grow poorly inestablished T cell lines. These isolâtes hâve been termed 25 M-tropic. The viral déterminant of T- and M- tropism mapsto alterations in the third variable région of gpl20 (theV3 loop) (Choe et al., 1996, Cell 85:1135-1148; Cheng-Mayeret al., 1991, J. Virol. 65 ; 6931-6941 ; Hwang et al., 1991,Science 253:71-74; Kim et al., 1995, J. Viral., 69:1755- 30 1761; and 0' Brien et al., 1990, Nature 348:69-73). The characterization of HIV- isolâtes with distinct tropismstaken together with the observation that binding to CD4cell surface protein alone is insufficient to lead toinfection, suggest that cell-type spécifie cofactors might 011307 be required in addition to CD4 for HIV-i entry into thehost cell.

2.2 TREATMENT FOR HIV INFECTION HIV infection is pandémie and HIV-associated diseases represent a major world health problem. Althoughconsidérable effort is being put into the design ofeffective Therapeutics, currently no curative anti-retroviral drugs against AIDS exist. In attempts todevelop such drugs, several stages of the HIV life cyclehâve been considered as targets for therapeuticintervention (Mitsuya, H., et-al., 1991, FASSE J. 5:2369-2381) . Many viral targets for intervention with HIV lifecycle hâve been suggested, as the prevailing view is thatinterférence with a host cell protein would hâvedeleterious side effects. For example, virally encoded reverse transcriptase has been one focus of drug development. A number of reverse-transcriptase-targeted» » — drugs, including 2 , 3 -dideoxynucleoside analogs such as AZT, ddl, ddC, and d4T hâve been developed which hâve beenshown to been active against HIV (Mitsuya, H., et al.,1991, Science 249:1533-1544).

The new treatment régimes for HIV-1 show that acombination of anti-HIV compounds, which target reversetranscriptase (RT), such as azidothymidine (AZT),lamivudine (3TC), dideoxyinosine (ddl), dideoxycytidine(ddC) used in combination with an HIV-1 protease inhibitorhâve a far greater effect (2 to 3 logs- réduction) on viralload compared to AZT alone (about 1 log réduction) . Forexample, impressive results hâve recently bee'n obtainedwith a combination of AZT., ddl, 3TC and ritonavir (Perelson, A.S., et al., 1996, Science 15:1582-1586) .However, it is li.kely that long-term use of combinations ofthese Chemicals will lead to toxicity, especially to thebone marrow. Long-term cytotoxic therapy may also lead to 011307 suppression of CD8+ T cells, which are essential to thecontrai of HIV, via killer cell activity (Blazevic, V., etal., 1995, AIDS Res. Hum.Retroviruses 11:1335-1342) and bythe release of suppressive factors, notably the chemokines 5 Rantes, MIP-lœ and ΜΙΡ-Ιβ (Cocchi, F., et al., 1995,Science 270:1811-1815). Another major concert in long-termChemical anti-retroviral therapy is the development of HIVmutations with partial or complété résistance (Lange, J.M.,1995, AIDS Res. Hum Retroviruses 10 :S77-82) . It is thought 10 that such mutations may be an inévitable conséquence ofanti-viral therapy. The pattern of disappearance of wild-type virus and appearnace of mutant virus due to treatment,combined with coincidental décliné in CD4 + T cell numbersstrongly suggests that, at least with some compounds, the 15 appearance of viral mutants is a major underlying factor inthe failure of AIDS therapy.

Attempts are also being made to develop drugs .whichcan inhibit vira'l entry into the cell, the earliest stageof HIV infection. Here, the focus has thus far been on 20 CD4, the cell surface . receptor for HIV. Recombinant soluble- CD4, for example, has been shown to inhibitinfection of CD 4^ T cells by some HIV-1 strains (Smith,D.H., et al., 1987, Science 238:1704-1707) . Certainprimary HIV-1 isolâtes, hawever, are relatively less 25 sensitive to inhibition by recombinant CD4 (Daar, E., et al., 1990, Proc. Natl. Acad. Sci. USA 87 .-6574-6579) . Inaddition, recombinant· soluble CD4 clinical trials hâveproduced inconclusive results (Schooley, R., et al., 1990/Ann. Tnt. Mëd. 112:247-253 ,· Kahn, J".O., et al., 1990, Ann. 30 Int. Med. 112:254-261,- Yarchoan, R., et al., 1989, Proc.

Vth Int. Conf. on AIDS, p. 564, MCP 137).

The -late stages of HIV réplication, which involvecrucial virus-spécifie processing of .certain viral encodedproteins, hâve also been suggested as possible anti-HIV 011307 drug targets. Lafce stage processing is dépendent on theactivity of a viral protease, and drugs are being developedwhich inhibit this protease (Erickson, J., 1990 Science249:527-533) .

Recently, chemokines produced by CD8 + T cells hâvebeen implicated in suppression of HIV infection (Paul,W.E. , 1994, Cell 82.-.17T; Bolognesi, D.P., 1993, Semin.Immunol. 5:203).

The chemokines RANTES, MIP-lœ and MIP-1E, which aresecreted by CD8 T cellSj were shown to suppress HIV-1 p24antigen production in cells infected with HIV-1 or HIV-2isolâtes in vitro (Cocchi, F, et al., 1995, Science270:1811-1815) .

Thus, these and other chemokines may prove useful inthérapies for HIV infection. The clinical outcome,however, of ail these and other candidate drugs is still inquestion.

Attention is also Being given to the development ofvaccines for the treatment of HIV infection. The HIV-1envelope proteins (gplSO, gpl20, gp41) hâve been shown tobe the major antigens for anti-HlV antibodies présent inAIDS patients (Barin et al.·, 1985, Science 228·. 1094-1096) .Thus far, therefore, these proteins seem to be the mostpromising candidates to act as antigens for anti-HIVvaccine development. Several groups hâve begun to usevarious portions of gpl60, gpl20, and/or gp41 as immunogenic targets for the host immune System. See forexample, Ivanoff, L., et al., U.S. Pat. No. 5,141,857;Saith, G., et al.,. WO92/22,654; Shafferman, A.,WO91/09,872 ; Formoso, C., et al·., WO90/Q7,119. To thisend, vaccines direct against HIV proteins are problematicin that the virus mutâtes rapidly rendering many of thesevaccines ineffective. Clinical results concerning thesecandidate vaccines, however, still remain far in thefuture. 011307 7

Thus although a great deal of effort is being directedto the design and testing o£ anti-retroviral drugs,effective, non-toxic treatments are still needed.

2. WASTING SYDROMSS

Wasting syndrome is a serious clinical problemcharacterized by a decrease in body mass of more than 10%from baseline body weight and a disproportionate loss ofbody mass with respect to body fat (Weinroth et al., 1995,Infectïous Agents and Disease Kotler and Grunfeld,1995, AIDS Clin. Rev. 96:229-275). Thus, wasting isdistinguished from starvation in which higher levais ofbody fat than body cell mass are depleted (Kotler et al.,1985r Am J. Clin. Nutr. 42:1255-1265; Cahill, 1970, N.Engl. J. Med. 282 -.668-675) . Wasting is associated with avariety of conditions, including HIV infection, otherinfectiou-s diseases^ sepsis, cancer, chronic ca-rdiovasculardisease and" diarrhea (Kotler et al., 1989, Am. J. Clin.Nutr. 50.:444-447,- Heymsfield et al., 1982, Am. J. Clin.Nutr. 36 :680-690) . Importantly, wasting is a significant factor in the mortality of patients -suffering frominfections or cancer. In fact, body cell mass déplétionhas a linear relationship to time of survival in AIDSpatients (Kotler et al., 1989, Am. J. Clin. Nutr. 50:444-447) .

The cause of wasting syndrome in AIDS and otherconditions is unclear and is most likely multifactorial.Metabolic abnormalities, irregular levels of ^hormones andcytokines, and malabsorption hâve ail been implicated inwasting syndrome. Not ail AIDS patients suffer fromwasting, suggesting that the cause of the wasting is notHIV itself. Most cases of HIV associated wasting syndromeare apparently caused by complications of AIDS, such assecondary infections and gastrointestinal disease (Kotlerand Grunfeld, 1995, AIDS Clin. Rev. 96:229-275) . 011307

Current and potential thérapies for wasting syndromesinclude nutritional support, appetite enhancers such asdronahinol and megestrol acetate, anabolic thérapies, suchas growth hormone, and cytokine inhibitors. However, mixedresults hâve been obtained with nutritional support andappetite enhancers in that patients tended to gain only fatand not overall body mass.. Administration of growthhormone, and cytokine inhibitors are still being tested andmay pose a risk of side effects (Kotler and Grunfeld, 1995,AIDS Clin. Rev. 96:229-275; Weinroth et al., 1995Infections Agents and Disease 4 :76-94).

Thus, treatment of wasting is critical to the survivaland well-being of patients suffering from serious diseasessuch as cancer and AIDS; thus,,- there is a need for .safe and.effective thérapies for wasting syndrome associated withcancer, AIDS and other infectious diseases.

2.4 PROPERTIES OF DIMETHYLFORMAMIDE AND OTHER POLAR

COMPOUNDS N,N' -Dimethylformamide (DMF) (molecular formula : C3H7ON)is a colourless, polar, hygroscopic liquid with lowvolatility and a boiling point of 152.5-153.5°C. It- isfreely miscible with water, alcohols and some hydrocarbons.DMF is generally used as a polar solvent and is readilyabsorbed through the skin, by inhalation, and upon oralingestion. DMF is rapidly metabolized, mainly in theliver, and excrétion accurs principally in the urine. Inrat, mouse, hamster and man the main métabolites of DMF are-N—hydroxymethyl-N-methylf ormamide (HMMF) , N-methylformamide (NMF) , and N-.acetyl-S- (N-methylcarbamoyl) cysteine (AMCC) ,as well as dihydroxymethylf ormamide (DHMF) and N-hydroxymethylformamide (HMF) . Unchanged DMF is ’excreted inthe urine as a small fraction of an administered dose of DMF. 011307 DMF has low acute dermal, oral and inhalationtoxicity. It is considered ta be a mild to moderate skinand eye irritant and readily permeates the skin. There isno indication of skin sensitizing properties. The 5 principal toxic effect of DMF and its métabolites is on theliver; DMF is well known to cause réversible hepatic damageassociated with typical clinical complaints, classicalbiochemical changes in the blood, and the appearance ofhépatocyte necrosis in liver biopsies. DMF is tératogénie, 10 but is not thought to be a mutagen or a carcinogen.

Viza et al·: hâve reported that DMF and DMSO inhibit in vitro réplication of HIV and Human Herpes Virus 6 (HHV-6)in certain cultured cell Unes. (See Viza et al., 1990AIDS Res. Hum. Retroviruses &xl31-132; Viza et al., 1989, 15 AIDS-FORSCHUNG 4:349-352; Viza et al., 1992, Antiviral Res. 18:27-38 and erratum at 19:179). DMF has been described as an in vitro differentiatingagent for certain transformed cells in culture- (SeeKoeffler, 1983, Blood 62:709-721; Calabresi et âl., 1979, 20 Biochem. Pharmacol. 28:1933-1941). when added to certainmalignant cells in vitro, DMF has- been reported to reducetheir tumorigenicity upori subséquent inoculation into nudemice (See Dexter, 1977, Cancer Res. 37:3136-3140; Dexter etal., 1979, Cancer Res. 39:1020-1025·). Upon intraperitoneal 25 injection into nude mince, DMF and NMF hâve been reportedto slow the growth of certain human cancer xenografts (SeeVan Dongen et al., 1989, Int. J. Cancer 43:285-292;Braakhuis et al., 1989, Head & Neck 11:511-515; Van Dongenet al., 1988, Acta Otolaryngol. 10-5:488-493, Dexter et al., 30 1982V Cancer Res. 42:5018-5Ô2~2) . However, the toxic side- effects of formamide and its N-methyl dérivatives in amouse sarcoma allograft model led investigators to concludethat these agents were unlikely to prove therapeuticallyuseful (See Clarke et al., 1953, Proc. Sec. Exp. Biol. Med. 35 84:203-207) . 011307 10

Attempts at treating human cancer patients with DMSOled to the conclusion that no objective response had beenshown (See Spremulli & Dexter, 1984, J. Clin. Oncol. 2:227-241) . Oral administration of NMF to human patients with 5 cancer- of the head and neck was reported as resulting inhepatotoxicity with no bénéficiai response (see Vogel etal., 1987, Invest. New Drugs 5:203-206), or with onlyminimal activity (See Planting et al., 1987, Cancer TreatRep. 71:1293-1294). 10 U.S. Patent No. 3,551,154 discloses the use of DMF as a pénétration enhancer to promote transdermal absorption oftopically applied médications. U.S. Patent No. 4,855,294discloses the use of glycerin to mitigate the skinirritation arising from the use of -DMSO and DMF as 15 pénétration enhancers to promote transdermal absorption oftopically applied médications. The lise of DMSO as apénétration enhancer 'to promote transdermal absorption ofantiviral agents is discussed in Woodford & Barry, 1986, J.Toxicol. Cut. & Ocular Toxicol. 5:167-177. 20 Citation or identification of any reference in Section 2 (or any other section) of this application shall not beconstrued as an admission that such reference is availableas prior art to the présent invention.

3 . SUMMARY OF THS INVENTION 25 The présent invention is directed to methods, compositions and drug delivery devices for treating ananimal affected by a viral or other microbial infection,especially an infection with a retrovirus such as HIV. Theinvention also provides methods, compositions and drug 30 delivery devices for treating an animal affected by awasting syndrome, such as wasting- associated with HIVinfection or malignancy.

According to the présent invention, there isadministered to an animal in need of treatment a 11 composition comprising N,N'dimethylformamide(dimethylformamide, DMF) ; N-hydroxymethyl N-methylformamide(HMMF) ; N-hydroxymethylformamide (HMF) ·dihydroxymethylformamide (DHMF); N-acetyl, S-(N-methylcarbamoyl)cysteine (AMCC); N-methylformamide (NMF);dimethylsufoxide (DMSO) ; formamide; acetamide,·methylacetamide, dimethylacetamide,· diethylacetamide;isopropylacetamide; diisopropylacetamide; N-acetylpiperidine; N-(β-hydroxyethyl)acetamide; N,N'-di(fi-hydroxyethyl)acetamide; N-acetylmorpholine, acrylamide;prepionamide; N-£luoromethyl-N-methyl-formamide,- pyridine-N-oxide; or any agent selected from the group consisting ofamides of the general formula Eg-CO-NRjR^, in which Rj andR2 are independen-tly selected’from the group consisting o-fH, methyl, halomethyl, saturated and unsaturated C2-C3 alkylgroups, and hydroxylated alkyl groups; or R| and R2 are together . selected from the groupconsisting of (0^)4, ((^2)5, and (-(¾) 20 (1^¾) 2; and R3 is selected from the group consisting of H, methyl,and saturated and unsaturated C2-C3 alkyl groups. Thetherapeutic composition may comprise a mixture of any twoor more of the aforementioned compounds.

In a patient infected with HIV, the therapeuticregimen may optionally combine a composition of the présentinvention with one or more additional agents effective fortreating HIV infection, including but not limited to agentsselected from the group consisting of nucleoside analogreverse transcriptase inhibitors, non-nucleoside reversetranscriptase inhibitors, and protease inhibitors, in anydesired combination.

The invention also extends to a vaccine prepared 'from antibodies that are obtained from the body of an animal after treatment with a composition of the invention for a viral infection. 12 011307

4 . BRIEF DESCRIPTION OF THS FIGTJRES

The présent invention may he more fully understood by reference to the following detailed description of theinvention, examples of spécifie embodiments of the S invention and the appended figures in which: FIGURE 1 illustrâtes plasma concentrations of DMF as a function of time in 4 patients treated with 2 DMF dermalpatches for 8 hours. FIGURE 2 illustrâtes plasma concentrations of DMF as10 a function of time in 3 patxents treated with 2 DMF dermal patches for 6 hours. FIGURE 3 illustrâtes HIV-1 viral load as measured byquantitative PCR in a patient treated with transdermal DMF.Two DMF patches were placed against the skin of the forearm 15 for 12 hours on days 0, 8, and 13 (indicated by arrows) . FIGURE 4 illustrâtes the general condition of HIVinfected patients before and after treatment withtransdermal DMF, as assessed according to the Karnofskyperformance scale. See text, Section 7, far details.

20 5. DETAILED DESCRIPTION OF THE INVENTION

The présent invention provides methods, compositionsand drug delivery devices for treating viral and microbialinfections. In one embodiment, the infection to be treatedis an infection with a retrovirus such as HIV, including an 25 asymptomatic infection, a latent infection, an infection accompanied by one or more symptoms of the AIDS-relatedcomplex, and an infection accompanied by clinical AIDS.Alternatively, the infection to be treated is any otherviral or microbial infection, including infection with 3'0- rubella, a herpesvirus such as Human Herpes Virus 6, theEpstein-Barr virus or cytomégalovirus, infection with anyvirus having a capsid protective coating, and anyopportunistic infection associated with disease of the 13 011307 immune System, such as an opportunistic infection in apatient infected with HIV.

The présent invention also provides . methods,compositions and drug delivery devices for treatment orprévention of any disease or disorder characterized by aloss of body mass. Particular conditions that can betreated by the methods and compositions of the - inventioninclude, but are not limited to, wasting associated withviral (e.g. HIV), bacterial, or any other types ofinfection or sepsis, cachexia associated with malignancy,chemotherapy or radiation therapy, wasting associated withchronic cardiovascular disease, wasting caused by exposureto toxic or radioactive substances, and wasting associatedwith diarrhea and other gastrôintestinal disordérs.

The subject to be treated may be-any animal,-includingbut not limited to a monkey, cow, sheep, ox, pig, horse,cat, dog chicken and the like, and is preferably. a mammal,more preferab-ly a primate, and most preferably a humanadult or child, for -instance a human child weighing at-least 3 kg. As used herein, the term "patient"' refers toany animal in need of treatment according to -the methods orcompositions of the présent invention.

According to the présent invention, there isadministered a therapeutic composition comprising DMF;HMMF; HMF; DHMF; AMCC; NMF; DMSO; formamidé; acetamide,·methylacetamide; dimethylacetamide; diethylacetamide;isopropylacetamide; diisopropylacetamide; N-acetylpiperidine, N-(S-hydroxyethyl) acetamide; N,N'-di (E-hydroxyethyl) acetamide; N-acetylmorpholine; acrylamide;p r op i onamide ,-N - f luo r orne t hyl - N -me thy 1 - formamidé ;pyridine-N-oxide; any agent selected from the group consisting ofamides of the general formula R^-CQ-NRjR^, .in whi'ch R| and S-2 are independently selected from the group consisting of H, methyl, halomethyl, saturated and 14 01 1 307 unsaturated C2-C3 alkyl groups, and hydroxylated alkylgroups; and R3 is selected from the group consisting of H, methyl,and saturated and unsaturated C2~c3 groups; or any agent selected from the group consisting of amides of thegeneral formula R^-CO-NR^R^, in which R| and R2 are independently selected from the groupconsisting of H, methyl, halomethyl, saturated. andunsaturated C2-C2 alkyl groups, and hydroxylated alkylgroups,· or R| and R2 are together selected from the groupconsisting of (0^)4, (CH2)j, and (CH2) 2O (CH2) 2; and R3 is selected from the group consisting of H, methyl,and saturated and unsaturated C2 -C3 alkyl groups.

In one spécifie emhodiment, at least one of Rj and R2 is amethyl group. In another spécifie emhodiment, at least oneof R| and R2 is' a fluorinated. C| - C3 alkyl group. The'therapeutic composition may comprise a mixture of any twoor more of the aforementioned compounds. Especiallypreferred is a composition comprising DMF.

For treatme'nt of an animal infected with HIV, thetherapeutic regimen may optionally include, in addition toa composition of the présent invention, one or more otheragents effective for treating HIV infection, for instanceone or more nucleoside analog reverse transcriptaseinhibitors such as zidovudine (AZT, ZDV), zalcitabine(ddC), didanosine (ddl), lamivudine (3TC), stavudine (d4T)?one or more-non-nucleoside reverse.transcriptase inhibitorssuch as nevirapine, delavirdine, loviride, atevirdine,pyridinone; one or more protease inhibitors .such assaquinavir, indinavir, ritonavir, nelfinavir; or any combination of the aforesaid or other anti-HIV therapeutic agents. The composition of the présent invention and the 15 011307 additional anti-HIV therapeutic agent or agents may beadministered simultaneously, sequentially, or in cycles oftreatment according to any desired therapeutic protocol.

The compositions of the présent invention may beadministered by any desired enterai or parentéral route,including but not limited to transderraal, intradermal,subcutaneous, intramuscular, intraperitoneal, intravenous,intranasal, épidural, intralymphatic and oral routes, Thecompounds may be administered by any convenient method, forexample by infusion or bolus injection, by absorptionthrough épithélial· or mucocutaneous linings (e.g., oral,gastric, intestinal or rectal mucosa, etc.) and may beadministered together with other biologically activeagents.

Administration can be systemic or local. In addition, itmay be désirable to introduce the pharmaceuticalcompositions of the invention into the central nervousSystem by any suitable route, including intraventricu-larand intrathecal injection; intraventricular injection maybe facilitated by an intraventricular cathéter, forexample, attached to a réservoir,- such as an Ommayaréservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and theformulation may include an aerosolizing agent. If desired,any two or more routes of administration may be employedsimultaneously, sequentially, or in cycles of treatmentaccording to any therapeutic protocol.

In a spécifie embadiment, it may be désirable toadminister a composition of the invention locally to thearea irr need of treatment; this may be achieved, -forexample and not by way of limitation, by topicalapplication, by injection, by means of a cathéter, bymeans of a suppository, or by means of an implant, saidimplant being of a porous, non-porous, or gelatinous le 011307 material, including membranes, such as sialastic membranes,or fibres.

In another embodiment, a composition of the inventioncan be delivered in a vesicle, in particular a liposome(see Langer, Science 249:1527-1533 (1990); treat et al., inLiposomes in the Therapy of Infections Disease and Cancer,Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327,- seegenerally ibid.)

In yet another embodiment, a composition of theinvention can be delivered in a controlled release System.

In one embodiment, a pump may be used- (see Langer,supra;- Sefton, CRC Crit. Ref. Biomed. Eng. 14.:201 (1987);Buchwald et al., Surgery 88:507 (1980); Saudek et al, N.Engl. J. -Med. 321:547 (1989) ) ._ In another embodiment,polymeric materials can be used (see Medical Applicationsof Controlled Release, Langer and Wise (eds.), CRC Près.,Boca Raton, Tlorida (1974); Controlled DrugBioavailability, _Drug Product Design and Performance,Smolen and Bail (eds.) Wiley, New York (1934J ; Ranger andPeppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:81 (1983): see also Levy et al., Science 225 .-190 (1985);During et al., Ann. Neurol. 25:351 (1989); Howard et al·.,J. Neurosurg. 71:105 (1989)). In yet another embodiment,a controlled release System can be placed in proximity ofthe therapeutic target, thus requiring only a fraction ofthe systemic dose (see, e.g., Goodson, in MedicalApplications of Controlled Release, supra, vol. 2, pp. 115-'138 (1984)).

Other controlled release Systems are discussed in thereview by Langer (Science 249 .-1527-1533 (1990) ) .

The présent invention also provides jpharmaceuticalpréparations. Such préparations comprise a therapeuticallyeffective amount of a composition of the invention and apharmaceutically acceptable carrier. In a spécifie 17 011307 embodiment, the terrn "pharmaceutically acceptable" meansapproved by a regulatory agency of the Fédéral or a Stateor other government or listed in the U.S. Pharmacopoeia orother generally recognized pharmacopoeia for use in5 animais, and more particularly in humans. The terrn"carrier" refers to a diluent, adjuvant, excipient, orvehicle with which the composition of the présent inventionis administered. Such pharmaceutical carriers can bestérile liquids, such as water and oils, including those of10 petroleum, animal, vegetable or synthetic origin, such aspeanut oil, soybean oil, minerai oil, sesame oil and thelike. Wateriis a preferred carrier when the pharmaceuticalpréparation is administered intravenously. Saline solutions and-aqueous-dextrose and glycerol solutions can15 also be employed as liquid" carriers, particularly forinjectable solutions. Suitable pharmaceutical excipientsinclude starch, glucose, lactose, sucrose, gelatin, malt,rice, flour, chalk,. silica gSl, sodium stéarate, glycerolmonostearate, talc, sodium chloride, dried skim milk,20 glycerol, propylene glycol, water, éthanol and the like.

The pharmaceutical préparation, if desired, can alsocontain minor amounts of wetting or emulsifying agents, orpH buffering agents. These préparations can take the formof solutions, suspensions, émulsion, tablets, pills,25 capsules, powders, sustained-release formulations and thelike. The préparation can be farmulated as a suppository,with traditional binders and carriers such as triglycérides. Oral formulation can includecarriers such as pharmaceutical grades o 30 lactose, starch, magnésium stéarate, sodium saccharine,cellulose, magnésium carbonate, etc. Examples of suitablepharmaceutical carriers are described in "Reminton'sPharmaceutical Sciences" by E.W. Martin. Such préparationswill contain a therapeutically effective amount of the 35 therapeutic, preferably in purified form, together with a standard mannitol. K 011307 suitable amount of carrier so as to provide the form forproper administration to the patient. The formulationshould suit the mode of administration.

The compositions of the présent invention may beadministered transdermally. In one embodiment, acomposition of the invention is directly applied to theskin. In another embodiment, a composition of theinvention is applied to a réservoir (e.g. a cotton woolpad, a synthetic polymer such as Teflon™, or any suitableadsorbant) that is applied to the skin, preferably under anocclusive dressing. In a spécifie embodiment, acomposition of the présent invention is applied to the skinby means of a dermal patch. The concentration of the activetherapeutic agent in the composition that is applied to theskin, contained in or adsorbed onto the réservoir, or contained in the dermal patch may be about 10-100%,preferably at least 50%, more preferably at least 90%.

In a preferred embodiment, a polar compound such asDMF is administered transdermally using any suitable drugdelivery device, for example by applying one or more dermalpatc’nes. The dermal patch may optionally comprise abacking, a réservoir such as an adsorbant impregnated witha polar compound of the invention, and a permeable membranethat is placed in contact with the skin. The backing maybe of any material, such as a natural or synthetic polymer,that resists Chemical attack by the polar compound.Especially preferred is a backing of high densitypolyethylene. The adsorbant may be a colloïdal substance,for instance diatomaceous earth or colloidal Silicondioxide. The permeable membrane may be of any- material that chemically resists the polar compound and may optionally be provided with pores. In a preferred embodiment, the permeable membrane is a Teflon^^ membrane having pores with a diameter of about 0.1 /im, a diameter of about 0.5 /xm, or a diameter within the range of fram about 011307 0.1 μτα to about 0.5 μτα. The patch may be self-adhesive ormay be held in contact with the skin by an applicator, suchas a wrapping or bandage, including without limitation anelastic bandage or an adhesive bandage; an Elastoplast TMbandage is suitable for this purpose. Preferably, thepatch contains a greater quantity of polar compound than isintended to be delivered through the skin of the patient tobe treated; for instance, the patch may contain about 50%more of the polar compound than is intended to bedelivered. The patch may be of any desired size and shape,and may for instance take the form of a disk approximately9cm in diameter.

In one embodiment, the patch comprises a polarcompound, such as DMF, and at least one otherpharmacologically active agent, for instance an local anti-irritant such as glycérine. In another embodiment, thepatch' comprises a polar compound, such as DMF, and has noothér pharmacologically active agent présent. In a furtherembodiment, the' patch comprises a polar compound, such asDMF, and has présent no other pharmacologically activeagent that is capable of being systemically absorbedthrough the skin,- or has présent no other pharmacologicallyactive agent in an mount that is systemically effectiveafter transdermal absorption. In yet another embodiment,the patch comprises a polar compound, such as DMF, and hasprésent no other systemically active pharmacological agent.

In yet a further embodiment, the patch comprises a polarcompound, such as DMF, and has présent no other antiviralagent, for instance acyclovir or arildone.

The présent invention thus provides a .dermal patchcomprising a polar compound, such as DMF, in an amount thatis effective for treating a viral or microbial infection(for example an infection with HIV) in a human adult or ahuman child. The présent. invention further provides adermal patch comprising a polar compound, such as DMF, in 20 011307 an mount that is effective for treating a wasting syndromein a human adult or a human child. In one embodiment, thepatch contains at least 0.25g of a polar compound such asDMF, preferably at least 0.5g, more preferably at least 1g,and still more preferably at least 5g, for instance 5-15gof a polar compound such as DMF.

To minimize evaporative loss of the polar compound,the patch may optionally be stored in a sealed container,such as a sealed polymer bag. If desired, each patch maybe individually sealed for convenience of use. Optionally,the patches may be refrigerated prior to use, for instanceat about 4°C, so as to reduce evaporative loss of the polarcompound. Preferably, the patches are prepared within 24hours of use and-are stored in.a sealed container at 4°C;however, patches are stable for one or more 'weeks whenstored in a sealed container at 4°C. Prior to applying apatch, an area of skin is preferably washed with a mild soap and water, rinsed fo remove soap residue, thoroughlydried, and then hydrated with a suitable skin lubricant andmoistening agent, such as jelly. The patch is thenremoved from the packing material and applied so that thepermeable membrane contacts the prepared skin surface. Thepatch may be held in place with an applicator. After adesired dosage period, the patch is removed. After removalof the patch, the treated area may be thoroughly washedwith a mild soap and water to remove any residue of thepolar compound.

The compositions of the présent invention may be~administered at any des-ired interval, for instance once,every two or three weeks,· once, twice or three times perweek; every second day; or daily. Preferably^ a.composition of the invention is administered in a dose' thatresults in a peak plasma level of about 2-200 mg/1, morepreferably about 100-200 mg/1, still more preferably about150 mg/1 of the active ingrédient of the composition, such 21 011307 as DMF. Especially preferred is a peak plasma level of100-150 mg/1 or 150-200 mg/1 of DMF. As used herein, theterm "ppm" refers to parts per million by weight, and is inpractice équivalent to mg/1. 5 For transdermal administration of a polar compound, the rate of absorption is determined by the skin of thesubject. Upon exposure to the human skin, liquid DMF isabsorbed at a steady-state rate of approximately 9.4mg/cm^/hour (see Mraz and Nohôva, 1992, Occup. Env. Health 10 64:85-92) . Accordingly, the desired rate of absorption may be achieved by controlling the surface area of the skin exposed to the drug, as by determining the area of each patch and the number of patches applied to the skin. For example, two patches of diameter 9 cm will expose a totalπ 15 skin surface area of 127 cnr to the polar compound; for DMF, this will resuit in an absorption rate of about 1.2 gof DMF per hour. An initial dose of about 15 mg/kg of DMTis especially preferred.

In one embodiment, a patient weighing about 72"kg is 20 initially treated with DMF by administering two 9 cmdiameter dermal patches for one hour, resulting in aninitial adsorbed dose of about 1.2 g of DMF, équivalent toabout 16.7 mg/kg. The number of patches may be scaled upor down, and a longer or shorter initial period of exposure 25 used, according to the body weight of the patient beingtreated. This starting dose may be repeated at any desiredinterval, as described above, and preferably is given atweekly intervals. Preferably, the patient is monitored forat least 72 hours after administration of DMF for signs of ' 3 0 toxicity, such as liver toxicity, for example by monitoring the sérum or plasma levels of enzymes such as aspartateaminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transferase (yGT) and alkaline phosphatase,proteins such as albumin, or substances such as conjugated 35. or unconjugated bilirubin. Preferably, sérum or plasma 22 011307 levels of AST and ALT do not exceed five times the upperlimit of normal as determined by t'ne reference range of thelaboratory and population in question; more preferablysérum or plasma levels of AST and ALT do not exceed three 5 times the upper limit of normal, most preferably AST and ALT are not elevated above normal levels or above pre-treatment levels. The dose of DMF may be escalated byàpplying the patches for successively longer periods; inone embodiment the patches are applied for two hours, then 10 four hours, then six hours, and so on; in anotherembodiment, the period of exposure is successively doubled.Preferably, the patient is monitored prior to any doseescalation, in order to detect any signs of toxicity. Ifdesired, the dose may be escalated at weêkly intervals. 15 The dose may be escalated in this manner until the dose is calculated to be about 150 mg/kg/dose,· or until a desiredpeak plasma levai, for instance 100-150 or 150-200 mg/1 isachieved; or until the period of exposure is about 6 hoursor about S'hours. One should be cautious before increasing 20 The dose above 240 mg/kg/dose. 6. EXAMPLE: TREATMENT O.F HIV INFECTION BY TRANSDERMALADMINISTRATION OF N, N'-DIMETHYLFORMAMIDE (DMF)Patients infected with HIV-1 were treated with dimethylformamide (DMF) by the application of skin patches25 impregnated with a dimethylf ormamide gel to the patient's body. N-acetyl-cysteine-glutathione and/or essentialphospholipids were administered (orally or intravenously)to the patients at a dosage of 250-mig to 3 00 mg daily as aliver booster. Instead or in addition, glutamine may also 30 be administered to the patient as a liver booster.

Two skin patches were applied to different parts of the patient's body, for example to the forearm. Each skinpatch contained about ,7,06g of gel comprising DMF (92,5 %m/m) and colloïdal silicone dioxide (7,5 % m/m) . The gel 23 011307 served to prevent leakage of liquid DMF from the patches.The patches were manufactured at most 12 hours prîor to useas DMF evaporates rapidly. The intended level of DMF inthe patient's blood was 100 ppm. For a patient weighingabout 60 kg, an amount of about 14g over a period of 12hours is required to produce a level of lOOppm. Based onstudies by Marz and Nohova the surface area required toabsorb this amount in about 127,2 cm^. To obtain a bloodlevel of 100 ppm about 1,272 g of DMF must be absorbed perhour, thus each sticker requires about 7,064 g DMF todeliver the required amount, having a surface area of 6,36cm^ (each sticker) . DMF absorption rate is 9,4 mg/cm^/hour.Xn theory this treatment will, deliver 125 - 135 ppm, butdue to évaporation of the DMF, 100 ppm was obtained.Absorption capabilifey varies from patient to patient dépending on factors such as skin-type and skin thickness.To obtain the desired levels of DMF in the patients, plasmaDMF concentrations were monitored for each patient andtreatment adjusted accordingly depending on the DMF levelof each patient. Figure 1 shows the plasma DMF levels of4 patients treated with 2 dermal DMF patches for 8 hours..Figure 2 shows the plasma DMF levels of 3 patients treatedwith 2 dermal DMF patches for 6 hours.

The stickers were thus each loaded with about 7,064 gof the gel of DMF and silicone dioxide. Each patch wasapplied for a period of 12 hours, either once per week overa period of 12 weeks, or twice per week over a period of 6weeks.

Blood tests in-certain patients _indicate an increasein CD4 T-cell counts from 350 to 1000, and a rapidréduction of PCR- (Polymerase chain reaction) (viral load)of 120,000 to 500/ml within three weeks with as little asthree treatments. Figure 3 shows serial quantitative PCRmeasurements of HIV-1 viral load in a patient prior totreatment and following three treatments with 2 dermal DMF 24 011307 pa.tch.es for 12 hours. The PCR test conducted was the Rocheamplicor HIV monitor. A viral load of < 500/ml plasma isconsidered to be undetectable.

Some patients undergoing treatment had severe acné or 5 displayed German measles symptoms before treatment. Whentreated with DM? so that the DMF level in the patient'sblood was 5 0 - 10 0 ppm, the German measles symptoms and thesevere acné cleared up or disappeared within 7 days.

Prior to the treatment with dimethylformamide, a 10 comprehensive base-line clinical and psychological évaluation of the patient was conducted. The évaluationprovided baseline biochemical and hematological data on thepatient. Detailed virological serology (HIV-1) tests werealso conducted to_ détermine the patient's total body virus 15 count, and these tests were conducted on a weekly basis, or as per treatment.

The concentration of DMF in the patient's blood wasdetermined hourly during the period _of treatment. Anintravenous line was introduced each morning to take blood 2 0 samples and was kept open with an infusion of Normal Saline at a rate of 20ml/hour and daily monitoring of the activemétabolite AMCC (eg by 4-hourly urine sampling) derivedfrom the DMF was also conducted. Subséquent applicationsof DMF were adjustedin accordance with measured changes in 25 blood level DMF concentration resulting from changes inabsorption variables and daily full haematological andbiochemical profiles were conducted to detect any changesin liver function. Daily full clinical and psychologicalévaluations were also conducted. - 30 A daily virological serology workout to establish total body virus count and to monitor improvements in theimmune status of the patient (CD4- T-helper cells) . andprognostic factors were also conducted. The serologyworkout was based on p24 antigen and quantitative PCR or,optionally, by other methods. A weekly détermination of 35 25 011307 • CD4 counts and Beta-2-macroglobulin was also conductedspecifically to monitor improvements in the patient'simmune status and prognosis. Ail clinical and laboratorydata were fed into a centralized data System to facilitaterapid response to any detrimental change so as to curtail treatment to maximize clinical effect and minimize potential side effects. The tests included a) Sérum: Na, K, Cl, CO2, Urea, Urate, Créatinine, Ca, Mg, Phosphate, Total and . Conjugated Bilirubin; b) Hematology: Hemoglobin, Red cell count, Hematocrit, MCV, MCH, MCHC, RDW; c) Sérum Pratein Electrophoresis: Total Proteins, Albumin, Total Globulin, Alphal Globulin, Alpha2Globulin, Beta Globulin, Gamma Globulin; d) White cell analysis·. Différentiel white cellcount, Absolute Neutrophil, Lymphocyte, Monocyte,Eosinophil and Basophil counts; 20 e) Liver Enzymes: Alk. Phos, Gamma GT, ALT (SGPT), AST JSGOT), LDH; f) Other: Cell markers, PCR, Beta2-Mocroglobulin,p24 Antigen, C-Reactive - protein, CK-MB concentration,· 25 g) Blood analysis for DMF levels; h) Urine analysis for AMCC levels It appears as if DMF acts as at least one of a reverse 30 transcriptase inhibitor and a protease inhibitor. In vitrotests were conducted and it appears ' that the solventproperties of DMF dissolve the virus particles, e.g. thecapsid.

EXAMPLE: TREATMENT OF HIV INFECTION BY—TRANSDERMAL 26 011307 ADMINISTRATION QF N,Ν'-DIMETHYLFORMAMIDE (DMP) A pilot study was conducted to evaluate the efficacy of transdermal DMF in the treatment of patients infectedwith HIV. Informed consent was obtained from each patient.Séropositive status was verified by Western blot using acommercial· kit for detecting antibodies to p24 (AbbottDiagnostics) and the presence of HIV-1 was documented byquantitative PCR using a commercial kit (Roche Âmplicor). DMF as obtained commercially (Sigma-Aldrich) and inplasma samples was analysed by mass spectroscopy/ gaschromatography (GC/MS) , using a Varian '9600 gaschromatograph, OV 351 column, carbowax-PEG _ capillarycolumn, and Finnegan Mat ITS40 ion . trap detector.Operational parameters were _ as follows: GC températureprogram: 60°C_for 1 min. followed by a température gradientof 9.4°C/min. for 20 min. MS ionization method: électronimpact; mass range: 40-80 mass units; 1 scan/sec,· peakthreshold: 3 counts/sec; background mass·. 69 mass units. A stock solution of dimethylacetamide was used as an internai standard. Ail samples were extracted with anorganic solvent containing the internai standard, allowedto precipitate for 30 min. in a refrigerator, andcentrifuged at 3000g for 5 min. before transfer to GC vialsfor injection. Rétention time of the DMF peak relative tothe internai standard was 3.26 minutes; the calibrationcurve showed a corrélation of 0.98. Quantitation of DMFappeared linear up to 100 mg/1 and the lower limit ofdétection was estimate at 0.5 mg/1 based an a signal-to-background ratio of 3:1.

Dermal patches 9 cm in diameter were used within 12hours of manufacture. Each patch had a backing of highdensity polyethylene (0.245 g), a permeable membrane ofTeflon^M (pore size 0.2 pim; 0.258 g) for placement againstthe skin, and containing 0.573 g of colloidal Silicondioxide impregnated with 7.067 g DMF between the backing 27 011307 and the permeable membrane. The patches were visuallyinspected to verify the absence of leakage and were weighedon an analytical balance to verify less than 10% déviationfrom a total mass of 8.153 g. The patches were applied tothe skin of the forearm and secured with Elastoplast^Mbandages. Either one or two patches were used for theinitial dose, as determined from the predicted skintransfer rate of 9.4 mg/crr/hr and the patient's bodyweight. A staged dose of DMF was used, with two-hourly bloodand urine sampling to détermine peak plasma DMF levels.Patients were dinically monitored for any toxic side-effects and were kept under comprehensive biochemicalsurveillance while the dosage was increased as necessary toachieve a peak plasma DMF level of 100-120 ppm. Once thecorrect dosage was established, transdermal DMF was givenonce a week. Initial évaluation -included daily détermination of: a) vital signs and body weight; b) clinical checklist and Karnofsky score; a) full blood count and érythrocyte sédimentationrate,· b) sérum urea, créatinine, glucose, sodium,potassium, ALT, AST, alkyline phosphatase andtotal bilirubin; c) Coulter analysis of CD4+ and CD8+ counts andCD4/CD8 ratio,· d) Quantitative estimâtes of HIV-1 load by PCR(Roche Amplicor) and analysis of antibody to p24(Abbott); e) Urinalysis (Dipstix).

At each weekly visit, the patients were evaluated for adverse effects, virtually ail these tests were repeated,

and blood and urine were collected for measurement of DMF and its métabolites. 28 011307

Patient 1 [ADF] began the protocol in relatively goocLhealth, complaining principally of pains in the arms andlegs and inability to sleep, Two DMF patches wereadministered once a week, for an average exposure period of8 hours. The average weekly dose of DMF was 6.il g and theresulting peak plasma DMF level average 75 mg/1. After 9weeks, the patient's CD4+ T cell count had increased from140 to 640 cells/μΐ and the PCR-measured viral load haddecreased from 250,000 to 50,000 copies/ml. After 10 weeksthe patient's weight had increased from 81.9 to 96.0 kg,the patient was clinically well and no longer complained ofpain in the limbs.

Patient 2 [AM] began the protocol complaining of lossof strength, inability to sleep, pains in the arms andlegs, and. had herpes sores in the mouth. One DMF patch wasadministered per week, for an average exposure period of 8hours. The average weekly dose of DMF was 7.12 g and theresulting peak plasma DMF level averaged 125 m/f. After 9weeks, the patient's CD4+ T cell count has increased from460 to 720 cells/μΐ, the PCR-measured viral load haddecreased from 29,000 to 13,000 copies/m?, and thepatient's weight had increased from 58.4 to 63.0 kg. Thepatient's herpetic sores had resolved, the limb pains haddisappeared and the patient appeared clinically well.

Patient 3 [SM] began the protocol with overt clinicalAIDS, and complained of respiratory difficulty. Two DMFpatches were administered once a week, for an averageexposure period of 8 hours. The average weekly dose of DMFwas 8.97 g and the resulting_peak'plasma DMF level averaged121 mg/Z. After 7 weeks, the patient's CD4+ T cell counthad increased from 39 to 138 cells/μ?, the PCR-measuredviral load had decreased from 222,000 to 160,000 copies/mC anH the patient's weight had increased from 74.2 to 100 kg.The patient's appetite had improved, the respiratory 29 011307 difficulties had resolved, the patient appeared clinicallywell and started exercising again.

Patient 4 [EM] began the protocol with secondaryinfections (including herpes) , anémia, diarrhea and acné.Two DMF patches were administered once weekly, for anaverage exposure period of 8 hours. The average weeklydose of DMF was 7.33 g and the resulting peak plasma DMFlevel ave'raged 90 mg/Z . .After 18 weeks, the patient's CD4+ T cell count had increased from 249 to 450 cells/μ?, thePCR-meastCred viral load had decreased from 13,000 to4,000 copies/mf, and the patient's weight had increasedfrom 81.5 to 90.4 kg. The patient appeared clinically well .and had no active medical complaints.

Patient 5 [SV] began the'protocol, complaining of poorappetite,· forgetfulness, abdominal pain, and severe fatiguethat prompted him to consider selling his interest in hisbusiness: Two DMF patches were administered once weekly, for an average exposure period of 6 hours. The averageweekly dose of DMF was 3.75 g and the resulting peak plasmaDMF leveT average 67 mg/Z. After 5 weeks, the patient'sCD4+ 'T cell count had increased from 354 to 396 cells/μ?,the PCR-measured viral load had decreased from 156,000 _to13,000 copies/mi, and the patient's weight had increasedfrom 56.0 to 58.0 kg. The patient was clinically well, hadacquired his business partner's share, and was running thebusiness himself.

Patent 6 [W] began the protocol complaining of secondary infections (including herpes), ataxia, andnumhness of the left arm and left side o£ -the face. TwoDMF patches were administered weekly, for an. averageexposure period of 8 hours. The average weekly dose of DMFwas- 8.25" g and the resulting peak plasma DMF level averaged110 mg/£. After 19 weeks, the patient’s CD4+ cell counthad increased from 260 to 450 cells/gf, the PCR-measuredviral load had decreased from 120,000 to 24,000 copies/m£, 30 011307 and the patient's weight had increased from 75.4 to 84.6kg. The secondary infections had resolved and the patientappeared clinically well.

Patient 7 [AJF] began the protocol severely ill. The 5 patient's initial CD4+ T cell count was 29 cell/μ?, and the initial PCR-measured viral load was 1,156,000 copies/m?,Treatment was with 1 patch, for an average exposure periodof 4 hours. The average weekly dose of DMF was 4.60 g andthe resulting peak plasma DMF level averaged 100 mg/?. 10 After the first treatment, the patient's CD4+ T cell countdecreased to 14 cells/μ?. Treatment was given daily forfive days _and then continued at weekly intervals. After 9treatments, the patient"s CD4 + T cell count had increasedto 35 cells/μ?, the _PCR-meastfred viral load had decreased 15 to 9,000 copies/m?, and the“patient's weight had increased from 46.5 kg (at commencement of DMF therapy) to 49.0 kg.The patient felt well .and had returned to full-timeemployment.

Patient 8 [MS] began the protocol with sevsre herpetic 20 lésions in the lower back and genitalia. Two DMF patches were given once weekly, for an average exposure period of·8 hours. The average weekly dose of DMF was 6.24 g and theresulting peak plasma DMF level averaged 130 mg/£. After 8weeks, the patient's CD4+ T cell count had increased from 25 200 to 240 cells/μί, the PCR-measured viral load had decreased from 1, 200,000 to 250,000 copies/m?. Thepatient's weight had increased from 48.1 to 52.2 kg, andthe Herpes lésions had entirely resolved.

Two patients were excluded from the trial, one due to 30 alcohol abuse and one due to viral hepatitis B.

Most patients experienced mild local skin irritation at the area of application after removal of the patches; the skin at the application site had a maculopapular appearance, probably due to intense hydration under the 35 patch. In one case, there was slight blistering that 31 011307 resolved within 24 hours and that did not cause 'the patientsignificant discomfort. Most patients experienced mildtransient nausea, usually on day three after treatment,which gradualiy decreased during the treatment .protocol;one patient reported moderate transient nausea. Fourpatients showed transient élévation of liver enzymes, whichnever exceeded three times the upper limit of normal andwhich in most cases returned ta pre-trèatment levels priorto the next dose of DMF. Most instances of elevated liverenzymes were associated with at least one factor unrelatedto the treatment protocol (alcohol consumption, hepatitis,and previous anti-HIV therapy with other agents). virtually ail patients showed clinica'l improvementafter 2-3 weeks of treatment.- As-shown by Figure 4, everypatient showed improved general condition after treatmentwith DMF when assessed according to the Karnofskyperformance scale, in which a patient's general status isassigned a numerical value as follows: 100 = normal, nocomplaints; 90 = able to carry on normal activities, minorsigns or symptoms of disease; 80 = normal activity witheffort; 70 = cares for self, unable to carry on normalactivity or to do active, work; 60 =- requires occasionalassistance, but is able to care for most needs; 50 =requires considérable assistance and frequent medical care,·40 = disabled, requires spécial care,· 30 = severely disabled, hospitalization indicated although death notimminent; 20 = very sick, hospitalization and activesupportive treatment necessary,· 10 = moribund, fatalprocess progressing rapidly; 0 = dead. (See Kanofsky etal., 1984, Cancer 1:634-656). Improvement in neurologicalsymptoms and Herpes viral infections was remarkable.Additional anti-microbial therapv for secondary infectionswas rarely needed. Within the first 14 days of DMFtreatment, there'was marked improvement in general fatigueand in appetite. Ail the patients gained weight. Clinical 32 01130? improvemènt correlated well with disea.se status as assessedby viral load and CD4+ T cell count. For five out of eightpatients, the relative PCR-measured viral load could befitted to Gampertz curves; this analysis revealed an 88.8% 5 décliné in PCR-measured viral load after 42 days of DMFtreatment. For seven out of eight patients, the relativeCD4± T cell count could be fitted to Gcmpertz curves; thisanalysis showed a 73.4% increase in CD4+ T cell countsafter 42 days of DMF treatment. 10 The présent invention is not to be limited in scope by the exemplified embodiments, " which are intended asillustrations of single aspects of the invention. Indeed,various modifications of the invention in addition to thoseshown and described herein will become apparent to those 15 skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended tofal-1 within the scope of the appended daims.

Ail publications cited herein are incorporated byreference in their entirety. ΠΓΤΓ

Claims (25)

1. 33 01130? CLAIMS

   1. A drug delivery device for transdermaladministration of a therapeutic agent, comprising: a réservoir containing or having absorbed thereon atherapeutic composition comprising N-hydroxymethyl-N-methylformamide (HMMF), N-hydroxymethylformamide (HMF),dihydroxymethylformamide (DHMF), N-acetyl-S-(N-methylcarbamoyl) cysteine (AMCC), dimethylsuifoxide (DMSO)or pyridine-N-oxide.
2. 2. A drug delivery device for transdermaladministration of a therapeutic agent, comprising: a backing,* a permeable membrane suitable for placement againstthe skin of an animal to. be treated; and an adsorbant situated between the backing and thepermeable membrane, the adsorbant having adsorbèd thereona therapeutic compositioih-comprising N, N' -dimethylformamide(DMF) , N-hydroxymethyl-N-methylformamide (HMMF) , N-hydroxymethylformamide (HMF), dihydroxymethylformamide(DHMF), N-acetyl-S-(N-methylcarbamoyl) cysteine (AMCC), N-methylformamide (NMF) , dimethylsulfoxide (DMSO) , formamide,acetamide, methylacetamide, dimethylacetamide,diethylacetamide, isopropylacetamide, diisopropylacetamide,N-acetylpiperidine, N-(0-hydroxyethyl) acetamide, N, N'-di (β-hydroxyethyl) a.cetamide, N-acetylmorpholine,acrylamide, propionamide, N-fluoromethyl-N-methyl^ f ormamide, pyridine-N-oxide, or an amide~ of the generalformula R^-CO-NRjl^' which R| and R2 are- independently selected from the groupconsisting of H, methyl, halomethyl, saturated and 3“ 011307 unsaturated C2-C3 alkyl groups, and hydroxylated alkylgroups or R| and R2 are together selected from the groupconsisting of (¢^2)4, (¢112)5, ((3¾) 20 (^¾) 2; and 5 R3 is selected from the group consisting of H, methyl, and satnrated and unsaturated C2-C2 alkyl groups; the drugdelivery device adapted to be placed in contact with theskin of an animal to be treated.
3. 3. The drug delivery device according to Claim 2, in 10 which the therapeutic composition comprises DMF.
4. 4. The drug delivery device according to Claim 3, inwhich the therapeutic composition comprises-at least 0.25g of DMF.
5. 5. The drug delivery device according_to Claim 4, in 15 which the therapeutic composition comprises at least ,5 g of DMF.
6. 6 . The drug delivery device according to any one of Claims 3 to 5 inclusive, in which the therapeuticcomposition comprises DMF in an amount that is effective 20 for treating a viral or microbial infection or a wasting syndrome in a human adult or a human child.
7. 7 . The drug delivery device according to any one of Claims 3 to S inclusive, having présent nopharmacologically active agent other than DMF.
8. 8 . The drug delivery device according to any one of Claims 3 to 7 inclusive, having présent nopharmacologically active agent, other than DMF, that iscapable of being systemically absorbed through the skin. 35 011507 9· The. drug delivery device according to any one of Claims 3 to 8 inclusive, in which the adsorhant iscolloïdal Silicon dioxide and the permeable membrane is aTeflon™ membrane having pores of diameter from about 0.lgm 5 to about 0.5/xm.
9. 10. A method of treating a wasting syndrome comprising administering to an animal in need of treatmenta therapeutic composition comprising N,N' -dimethylformamide(DMF), N-hydroxymethyl N-methylformamide (HMMF), N- 19 hydroxymethylformamide (HMF) , dihydroxymethylformamide (DHMF), N-acetyl, S-(N-methylcarbamoyl) cysteine (AMCC), N-methylformamide (NMF), dimethylsulfoxide (DMSO), formamide,acetamide, methylacetamijde, , dimethy lace tamide^ diethylacetamide, isopropylacetamide, diisopropylacetamide, 15 N-acetylpiperidine, N-(β-hydroxyethyl) acetamide, N,N'-di(β-hydroxyethyl) acetamide, N-acetylmorpholine,acrylamide, propionamide, - N-fluoromethyl-N-methyl-formamide, pyridine-N-oxide, or an amide of the generalformula R3-CO-NR1R2, in which 20 R^ and R2 are independently selected from the group consisting of H, methyl, halomethyl, saturated - andunsaturated C2 - C3 alkyl groups, and hydroxylated alkylgroups; or Rj and R2 are together selected from the group 25 consisting of (CH2)4, (CH2)5, and (CH2)202' an<^ R3 is selected- from the group consisting of H, methyl, and saturated and unsaturated C2 - C3 alkyl groups.
10. 11. The method according to Claim 10, in which the therapeutic composition comprises an amide of’the generalformula R3-CO-NR2R2, in which 30 36 011307 R| and R2 are independently selected from the groupconsisting of H, methyl, halomethyl, saturated andunsaturated C2-C3 alkyl groups, and hydroxylated alkylgroups; or 5 Rj and R2 are together selected from the group consisting of (CH-p^, (CH^^, and (CH2) 2O (CH2) 2; and R3 is selected from the group consisting of H, methyl,and saturated and unsaturated C2-C2 alkyl groups.
11. 12. The method according.to Claim 10 or Claim 11, in 10 which the therapeutic composition comprises an amide of the general formula R^-CO-NRjR^ "in which R| and R2 are independently selected from the group consisting of H,· methyl, halomethyl, saturated andunsaturated C2-C3 alkyl groups, and hydroxylated alkyl 3.5 groups; and R^ is selected'from the group consisting of H, methyl,and saturated and, unsaturated C2-C2, alkyl groups, andhydroxylated alkyl groups.
12. 13 . The method according to any one of Claims 10 to 20 12 inclusive, in which the therapeutic composition comprises DMF, HMMF, HMF, DHMF, AMCC, NMF, DMSO, formamide,acetamide, methylacetamide , dimethylacetamide, diethylacetamide, isopropylacetamide, diisopropylacetamide,N-acetylpiperidine, N-(β-hydroxyethyl) acetamide, N,N'- 25 di(β-hydroxyethyl) acetamide, N-acetylmorpholine, acrylamide, propionamide, N-f_luaromethyl-N-methyl-formamide, or pyridine-N-oxide.
13. 14. The method according to Claim 13, in which the therapeutic composition comprises DMF, HMMF, HMF, DHMF, 30 AMCC, or NMF. TfV 37 011307
14. 15. The method according to any of Claims 10 to 14inclusive, further comprising administering at least oneother agent effective for treating HIV infection.
15. 16. The method according to Claim 15, furthercomprising administering at least one nucleoside analogreverse transcriptase inhihitor, at least one non-nucleoside reverse transcriptase inhihitor, or at least oneprotease inhihitor.
16. 17 . The method according to any one of daims 14 to16 inclusive, in which the therapeutic compositioncomprises DMF.
17. 18. The method according to any one of Claims 13, 15or 16, in which the therapeutic composition comprises DMSO.
18. 19. The method according to any one of Claims 10 to 18 inclusive, in which the therapeutic composition isadministered transdermally.
19. 20. A method of treating _a viral or microbialinfection, comprising administering to an animal in need oftreatment a therapeutic composition comprising N-hydroxymethyl N-methylformamide (HMMF), N-hydroxymethylformamide ' (HMF), dihydroxymethylformamide(DHMF), N-acetyl-S-N-methylcarbamoyl) cysteine (AMCC),dimethylsulfoxide (DMSO) or pyridine-N-oxide.
20. 21. A method as claimed in Claim 20, in which thetherapeutic compositions is administered for treatment ofa retroviral infection.
21. 22. A method as claimed in Claim 21, in which theretroviral infection is an HIV infection. 38 011307
22. 23. A method according to any one of Claims 20 to 22inclusive, further comprising administering at least oneother agent effective for treating HIV infection.
23. 24. A method as claimed in any one of Claims 20 to 23 5 inclusive, further comprising administering at least one nucleoside analog reverse transcriptas inihibitor, at leastone non-nucleoside reverse transcriptase inhibitor, or atleast one protease inhibitor.
24. 25. A method as claimed in any one of Claims 20 to 24 10 inclusive, in which this therapeutic agent. is administered transdermally.
25. 26. A vaccine prepared from antibodies harvested fromthe body of an animal treated for a viral infectionaccording to the method of Claim 20.