USOO5409690A United States Patent 19 11 Patent Number: 5,409,690 Howell et al. (45) Date of Patent: Apr. 25, 1995 54 TREATMENT OF MULTIDRUG RESISTANT Primary Examiner-Raymond Henley, III DISEASES IN CANCER CELL BY Assistant Examiner-T. J. Criares POTENTATING WITH MASOPROCOL Attorney, Agent, or Firm-Weiser & Associates 75) Inventors: Stephen Howell, Del Mar, Calif.; Atul Khandwala, Edgewater, N.J.; Om P. 57 ABSTRACT Sachdev, New City, N.Y.; Charles G. Disclosed are: a method for reversing multidrug resis Smith, Rancho Santa Fe, Calif. tance in a mammal; and a composition to reverse multi 73) Assignee: Chemex Pharmaceuticals, Inc., Fort drug resistance comprising: Lee, N.J. 21 Appl. No.: 81,663 is is , is R9 22 Filed: Jun. 23, 1993 R1O g--- 51) Int. Cl. ..................... A61K 48/00; A61K 31/70; R10 R11 R12 R13 R8 A61K 31/045 R2O R7 52 U.S. C. ........................................ 424/10; 514/34; 514/727; 514/731 wherein 58 Field of Search ......................... 514/34, 7. R1 and R2 are independently H, lower alkyl or lower acyl; (56) References Cited R3, R4 and R5 are independently H or lower alkyl; U.S. PATENT DOCUMENTS R7, R8 and R9 are independently H, hydroxy, lower alkoxy or lower acyloxy; and 5,008,294 4/1991 Jordan et al. ....................... 514/73 R10, R11,y R12 and R13yIOXy are independently H or lower OTHER PUBLICATIONS alkyl, in a pharmaceutically acceptable vehicle. Carter et al; Chemotherapy of Cancer, Second Edition, A Wiley Medical Publication, 1981 pp. 361-365. 22 Claims, No Drawings 5,409,690 1. 2 brew tea which was the basis for a folk remedy that TREATMENT OF MULTIDRUG RESISTANT called for drinking the tea to cure colds, rheumatism DISEASES IN CANCER CELL BY POTENTIATING and other ailments. However, this remedy has not WITH MASOPROCOL proven to be successful. A clinical study was conducted by Smart et al, re BACKGROUND OF THE INVENTION ported in U.S. Pat. No. 4,880,637, in which human can Field of the Invention cer patients ingested either a tea made form the creosote This invention relates to a method of preventing bush or doses of pure NDGA. This study indicated that development of multidrug resistance (hereinafter neither NDGA nor the tea were effective anticancer MDR) and to a method of reversing MDR if one al 10 agents and in some cases caused stimulation of tumor ready exists thereby to prevent or correct drug accumu cell growth. This confirmed the earlier screening stud lation defect in drug resistant cells. More particularly, ies of NDGA conducted by Leiter et all of the Cancer the invention relates to the administration of maso Chemotherapy National Service Center of National procol along with antineoplastic/cytotoxic drugs to Cancer Institute which obtained negative results when which cells develop multidrug resistance (MDR), e.g. 15 NDGA was tested against several types of cancer cells. Doxorubicin, Daunorubicin, Amsacrine Mitoxantrone, It has been reported in U.S. Pat. No. 4,880,637 that Dactinomycin, Ellipticine, Etoposide, Teniposide, NDGA and its analogues in combination with ionic zinc Chlorambucil, Melphalan, Cyclophosphamide, Ni is effective in the treatment of benign, paramalignant trosoureas BCNU, CCNU, MeCCNU), Methotrexate, and malignant growth of the skin without detrimental Trimetrexate, 5-FU, Ara-C, Ara-A, Cisplatin, Carbo 20 side effects associated with chemotherapy or chemosur platin and Taxol. gical techniques. Reported Developments NDGA was also reported in U.S. Pat. No. 4,695,590, Patients having solid malignant tumors e.g., breast, as having efficacy in retarding senescence or aging in ovarian, lung, colon and hematological malignant disor mammals. ders, such as refractory myeloma, often develop MDR 25 To our knowledge, NDGA in combination with . MDR is associated with the overexpression of an other cytotoxic/antineoplastic chemotherapeutic drugs MDR-1 gene that codes for a plasma membrane P glycoprotein. The expression of the MDR-1 gene is is not known, nor have been reported to inhibit multi believed to be associated with a decreased cellular accu drug resistance associated with the treatment of solid mulation of drug due to an active energy dependent 30 malignant tumors or hematological disorders in mam efflux mechanism. The active efflux of drug is believed mals. to be mediated by the P-glycoprotein efflux pump. In the modern clinical oncology practice, it has been Attempts have been made to reverse MDR and to established that overexpression of P-glycoprotein (Pgp) correct the drug accumulation defect in drug resistant in cancer patients is directly related to the increase in cells. Among the agents which show promise in vitro 35 the multidrug resistance (MDR) to treatment with pres are verapamil, quinidine, amioderone, cyclosporin and ently available chemotherapies. The present inventors certain phenothiazines. It appears, however, that in have discovered that masoprocol (NDGA) overcomes vivo concentrations necessary to reverse MDR cannot the Pgp mediated resistance and Pgp antagonism of the be achieved without substantial toxicity to patients. cells, thereby increasing the levels of chemotherapeutic drugs which accumulate in the cancer cells. SUMMARY OF THE INVENTION The present invention provides a method and compo Masoprocol is new MDR modulating agent. We have sition for the inhibition and/or reversal of multidrug discovered that the combination of masoprocol known resistant phenomenon in a patient and thereby treatment as nordihydroguaiaretic acid (hereinafter sometimes of solid malignant tumors and hematological malignan referred to as NDGA) or certain analogues thereof in 45 cies comprising the administration of NDGA to said combination with cytotoxic agents, e.g. Doxorubicin, patient preferably in a pharmaceutically acceptable Daunorubicin, Amsacrine, Mitoxantrone, Dactinomy vehicle. cin, Ellipticine, Etoposide, Teniposide, Chlorambucil, The general formula comprises of from about 0.1% to Melphalan, Cyclophosphamide, Nitrosoureas (BCNU, about 25% NDGA or an analogue thereof in a pharma CCNU, MeCCNU), Methotrexate, Trimetrexate, 5-FU, 50 ceutically acceptable vehicle for topical application or Ara-C, Ara-A, Cisplatin, Carboplatin and Taxol can of from about 1 mg NDGA to about 500 mg of NDGA overcome multidrug resistance of certain diseased cells per kg of body weight for systemic administration. and has the potential of being effective in the treatment NDGA and its analogues used in the present inven of solid malignant tumors e.g., brain, breast, colon, lung, tion have the following Formula (I): ovarian cancers and hematological malignant disorders 55 including lymphoma, leukemia (acute nonlymphocytic leukemia, acute myelocytic leukemia), or increase the is is , is /y-R. " therapeutic effectiveness of the above-mentioned cyto toxic agents. R1O-(U)---- The naturally occurring meso form of the nordihy R10R11 R12 R13 R8 droguaiaretic acid meso- 1,4-bis (3,4-dihydroxy R2O R7 phenyl)-2,3-dimethylbutane ("NDGA') was reported as providing a positive result against malignant mela wherein noma. C. R. Smart, et al, Rocky Mountain Medical Jour R1 and R2 are independently H, lower alkyl or lower nal, November 1970, pp.39-43. (Unless otherwise indi 65 acyl; cated, NDGA is used herein to refer to the meso form R3, R4, R5, and Ré are independently H or lower of nordihydroguaiaretic acid). NDGA is found in the alkyl; R7, R8 and R9 are independently H, hydroxy, creosote bush, and this plant was used for centuries to lower alkoxy or lower acyloxy; and 5,409,690 3 4 R10, R11, R12 and R13 are independently H or lower be adjusted by methods well known in the art, for exam alkyl. ple, by the use of a higher molecular weight polyethyl Lower alkyl is intended to generally mean C1-C6 ene glycol. alkyl, and preferably R3 and R4 are C1-C3 alkyl. Lower In addition to NDGA, the formulation can contain acyl is intended to generally mean C1-C6 acyl, with pharmacologically-acceptable additives or adjuvants C2-C6 acyl being preferred. The formula is directed to such as antimicrobial agents, e.g. methyl, ethyl, propyl both the phenolic compounds and the conventional and butyl esters of parahydroxybenzoic acid as well as esters and ethers thereof. chlorobutanol, phenol, ascorbic acid, etc. The formula tion can also contain thickening or gelling agents, emul DETAILED DESCRIPTION OF THE 10 sifiers, wetting agents, coloring agents, buffers, stabiliz INVENTION ers and preservatives including antioxidants such as NDGA and its analogues (also referred to herein as butylhydroxyanisole in accordance with the practice of catecholic butanes) according to the present invention, the art. The formulation can also contain penetration may be made by the synthetic methods provided in U.S. enhancers such as dimethylsulfoxide, long-chain alco Pat. Nos. 4,562,298 and 4,954,659 which are incorpo 15 hols such as nonoxynol, long-chain carboxylic acids, rated herein by reference in their entirety. propylene glycol, N-(2-hydroxyethyl)pyrrolidone, 1 Examples of catecholic butanes include: the d-, 1 dodecyl-azacycloheptan-2-one, and the like. Depending racemic mixture of d- and 1-, meso-isomers of 1,4- on the method of application and the disease being bis(3,4-dihydroxyphenyl)-2,3-dimethylbutane; 1,4- treated, it may be desirable to use absorption-delaying bis(3,4-dihydroxyphenyl)butane; 1,4-bis(3,4-dimethoxy 20 agents such as aluminum monostearate and gelatin. phenyl)-2,3-dimethylbutane; 1,4-bis(3,4-diethoxy The compositions of the formulation can be adjusted phenyl)-2,3-dimethylbutane; 1,4-bis(3,4-dipropoxy using components well-known in the formulation art to phenyl)-2,3-dimethylbutane; 1-(3,4-dihydroxyphenyl)- provide a pharmaceutical formulation which is a gel, 4-(3',4',5'-trihydroxyphenyl)butane; 1,4-bis(3,4- cream, ointment, solid, liquid, semi-solid, etc. The par diacetoxyphenyl)-2,3-dimethylbutane; 1,4-bis(3,4-dipro 25 ticular physical form of the formulation depends on the pionyloxyphenyl)2,3-dimethylbutane; 1,4-bis(3,4- desired method of treatment and the patient to be dibutyroyloxyphenyl)-2,3-dimethylbutane; 1,4-bis(3,4- treated.