WO2015131039A1 - Method of treating neoplastic epithelial lesions - Google Patents

Abstract

Disclosed is a method of treating a neoplastic lesion in epithelium of a mammalian subject includes administering locally to a site of the lesion a therapeutically effective amount of a composition including a non-allium anti-fungal agent. Also disclosed is a method of treating a neoplastic lesion in epithelium of a mammalian subject comprising administering systemically a therapeutically effective amount of a composition including a non-allium anti- fungal agent, wherein the anti- fungal agent is selected from the group consisting of an agent that inhibits squalene monoxygenase and an agent that inhibits 1,3-beta glucan synthase.

Description

Method of Treating Neoplastic Epithelial Lesions

Cross Reference to Related Applications

[0001] This Application claims priority to U.S. provisional application serial number 61/946,341 filed February 28, 2014, which is hereby incorporated by reference in its entirety.

Field of the Invention

[0002] The present invention relates to methods of treating neoplastic epithelial lesions.

Background Art

[0003] Neoplastic epithelial lesions are common. Hyperkeratosis, one form of epithelial lesion, is understood as rendering less effective a treatment for fungal infections, such as those causing athlete's foot. See Goodman and Gilman, The Pharmacological Basis of Therapeutics, 1985, (the bible of therapeutics) p. 976.

Summary of the Embodiments

[0004] In a first embodiment of the invention there is provided a method of treating a neoplastic lesion in epithelium of a mammalian subject comprising administering locally to a site of the lesion a therapeutically effective amount of a composition including a non-allium anti- fungal agent. In some embodiments, the agent is a synthetic anti- fungal agent. In related embodiments, the neoplastic lesion is a carcinoma. Optionally, the carcinoma is a basal cell carcinoma or a squamous cell carcinoma. Optionally, the subject is a human.

[0005] In other related embodiments, the neoplastic lesion is a melanoma or a hyperkeratotic lesion. In other related embodiments, the neoplastic lesion is a nevus. [0006] In various embodiments, administering includes applying the composition topically to the lesion, or injecting the composition into the lesion, or injecting the composition subcutaneously.

[0007] In another related embodiment, the anti- fungal agent inhibits synthesis of ergosterol in a fungus. In a further related embodiment, the anti- fungal agent inhibits the enzyme squalene monoxygenase (also called squalene epoxidase). In another related embodiment, the anti- fungal agent inhibits the enzyme lanosterol 14 alpha-demethylase. In another related embodiment, the anti- fungal agent inhibits the enzyme 1 ,3-beta glucan synthase.

[0008] Optionally, the composition further comprises a chemo therapeutic agent. Also optionally, the composition further comprises a cosmetic agent (e.g., a pigment or dye).

[0009] In another embodiment of the invention there is provided method of treating a neoplastic lesion in epithelium of a mammalian subject comprising administering

systemically a therapeutically effective amount of a composition including a non-allium antifungal agent, wherein the anti-fungal agent is selected from the group consisting of an agent that inhibits squalene monoxygenase (also called squalene epoxidase) and an agent that inhibits 1 ,3-beta glucan synthase. In some embodiments, the agent is a synthetic anti- fungal agent. In related embodiments, the neoplastic lesion is a carcinoma. Optionally, the carcinoma is a basal cell carcinoma or a squamous cell carcinoma. Optionally, the subject is a human.

[0010] In other related embodiments, the neoplastic lesion is a melanoma or a hyperkeratotic lesion. In other related embodiments, the neoplastic lesion is a nevus.

[0011] In various embodiments, administering includes injecting the composition intravenously into the subject, injecting the composition intramuscularly into the subject, injecting the composition intrathecally into the subject, or injecting the composition intraperitoneally into the subject. In related embodiments, administering includes oral administration of the composition to the subject. Brief Description of the Drawings

[0012] The patent or application file contains at least one drawing executed in color. Copies of this patent with color drawing(s) will be provided by the Patent and Trademark Office upon request and payment of necessary fee.

[0013] The foregoing features of embodiments will be more readily understood by reference to the following detailed description, taken with reference to the accompanying drawings, in which:

[0014] Figs. 1 , 2, and 3 are a series of photographs of a basal cell carcinoma. Fig. 1. shows the basal cell carcinoma lesion before treatment; Fig. 2 shows the lesion after treatment had been in progress for 8 weeks; and Fig. 3 shows the lesion after treatment for 21 weeks.

[0015] Figs. 4, 5, and 6 are a series of photographs of a senile keratotic lesion. Fig. 4 shows the senile keratotic lesion taken before treatment. Fig. 5 shows the lesion after treatment had been in progress for 8 days, and Fig. 6 shows the lesion after treatment for 16 days.

[0016] Figs. 7, 8, and 9 are a series of photographs showing one example of a seborrhoeic keratosis (SK) located on the front of the leg. Fig. 7 shows the SK lesion taken on the day that treatment started, Fig. 8 is a photograph of the same lesion 7 days after treatment had begun, and Fig. 9 is a photograph of the same lesion after treatment for 15 days.

[0017] Figs. 10, 11 , and 12 are a series of photographs showing an actinic keratosis lesion. Fig. 10 shows the actinic keratosis lesion before treatment; Fig. 11 shows the lesion after treatment had been in progress for 4 weeks; and Fig. 12 shows the lesion after treatment for 6 weeks.

Detailed Description of Specific Embodiments

[0018] The use of anti-fungal agents to treat fungal infections is common. Antifungal agents, such as tolnaftate, are sufficiently old enough to be ignored in The Merck Manual (1999) and not listed in The Johns Hopkins Handbook of Drugs 1993. The

Merck Manual online, The Mayo Clinic's website and webMD.com list the only use of tolnaftate as a topical anti- fungal agent. The presence of hyperkeratosis, one form of epithelial lesion, at the site of application is described as rendering tolnaftate a less effective treatment for fungal infections, such as those causing athlete's foot. See Goodman and Gilman, The Pharmacological Basis of Therapeutics, 1985, (the bible of therapeutics) p. 976.

[0019] Despite these teachings of the art, the inventors have unexpectedly discovered that administration of an anti-fungal agent can be used to treat a neoplastic lesion in humans.

[0020] The publications, web sites, company names, and scientific literature referred to herein establish the knowledge that is available to those with skill in the art and are hereby incorporated by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. Any conflict between any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter.

[0021] Definitions. As used in this description and the accompanying claims, the following terms shall have the meanings indicated, unless the context otherwise requires:

[0022] An "anti-fungal agent" is an agent that prevents fungal growth when administered to a patient with a fungal infection.

[0023] A "neoplastic lesion" is a population of aberrant cells in epithelium of a mammal. In some cases, the neoplastic lesion can be observed from the body surface of the mammal. In some cases, the neoplastic lesion is a carcinoma (e.g., a basal cell carcinoma or a squamous cell carcinoma). In some cases, the neoplastic lesion is a melanoma. In some cases, the neoplastic lesion is a hyperkeratotic lesion. In some cases the neoplastic lesion is a mole or nevus.

[0024] To administer a composition "locally" to an epithelial lesion includes applying the composition topically at the site of the lesion, injecting the composition into the lesion, and/or injecting the composition at the site of the lesion (e.g., the composition may be injected subcutaneously at the site of the lesion). The composition may be in many forms including a cream, a liquid, an aerosol, a gel, or a powder. In instances wherein the epithelium is in the lining of the alimentary canal, for example, local administration may be oral. [0025] "Epithelium" of a mammal is a normally unbroken layer of densely adherent cells that cover a topologically external surface of the mammal. The epithelium includes, among other things, the skin and the lining of the alimentary canal, salivary gland, and ovary.

[0026] To "treat" a lesion means to reduce the growth of a lesion (e.g., slow the growth of the cells of lesion), or to inhibit the growth of a lesion (e.g., to slow or prevent an increase in size of the lesion), or to prevent the growth of a lesion (e.g., to prevent the start of growth of the lesion or prevent increased growth of cells of the lesion), or to shrink (e.g., ablate or induce regression of) the lesion.

[0027] A "synthetic agent" is a man-made agent that is not chemically identical to an agent that has been isolated from a naturally occurring organism.

[0028] A "non-allium" agent is an agent that is not chemically identical to an agent that is derivable from a plant in the genus Allium.

[0029] In accordance with protocols implementing embodiments of the present invention, fourteen neoplastic lesions have been treated with anti- fungal creams and a liquid gel. Initially, a smaller dose was applied daily. After a period of time, one lesion stopped growing but then stopped regressing, which was overcome by doubling the dose and frequency of application.

[0030] First protocol. Dosage: about 0.05 ml = 2mg was applied to the lesions once a day and left uncovered.

[0031] Second Protocol. Dosage: about 0.1 ml = 4mg was applied to the lesions twice a day and covered with a latex-free sterile band aid.

[0032] The antifungal agents were:

[0033] 1. Tolnaftate cream 1%, sold by MSD, a subsidiary of Merck that was not listed in the Mark Manual of 1999 indicating ignorance of its present therapeutic potential. It was also not listed in The Johns Hopkins Handbook of Drugs 1993. It is a thiocarbamate that somehow inhibits ergosterol synthesis. Ergosterol is to a fungus what cholesterol is to humans. Tolnaftate is a well-known anti- fungal agent having the IUPAC name 0-(2- naphthyl)methyl(3-methylphenyl)thiocarbamate. It has a somewhat selective spectrum of action excluding, for example, Candida. Tolnaftate is believed to inhibit the enzyme squalene monoxygenase (also called squalene epoxidase), although its exact mechanism of action is unknown. [0034] It was applied to seborrhoeic keratosis (lesion SK 1, Figs. 7-9), to basal cell carcinoma (lesion BCC 1 , Figs. 1-3), melanoma (lesion M l), and to a nevus lesion (lesion N

1).

[0035] 2. Clotrimazole cream 1%, sold by MSD, a subsidiary of Merck. It is an azole, it inhibits the enzyme lanosterol 14 alpha-demethylase which is a necessary step for the production of ergosterol which is part of the cell wall of fungi.

[0036] It was applied to actinic keratosis (lesions AK 2, Figs. 4-6; and AK 9, Figs. 10-12), and a second nevus (lesion N 2).

[0037] 3. Butenafine HCl cream 1%, sold by MSD, a subsidiary of Merck. It is an allylamine, it inhibits squalene monoxygenase (also called squalene epoxidase), which is a necessary step in the production of ergosterol which is part of the cell wall of fungi.

[0038] It was applied to actinic keratosis (lesions AK 4 and AK7).

[0039] 4. Terbinafine HCl cream 1%, sold by Novartis. It is an allylamine, it inhibits squalene monoxygenase (also called squalene epoxidase), which is a necessary step in the production of ergosterol which is part of the cell wall of fungi.

[0040] It was applied to actinic keratosis (lesion AK 3) and seborrhoeic keratosis (lesion SK 2).

[0041] 5. Undecylenic acid liquid gel 25% w/w, sold by Alva-Amco, a naturally occurring unsaturated fatty acid extracted from castor beans (Ricinus communis). It inhibits hyphae formation apparently by blocking fatty acid synthesis.

[0042] It was applied to actinic keratosis (lesions AK 1 and AK 5).

[0043] The lesions (N = 15), comprised:

[0044] Two seborrhoeic keratosis lesions: SK 1 & SK 2. Deeply melanotic seborrhoeic keratosis, the lesions grew over a two-three year period. The protuberant rough surface was very firmly attached. Diameter 1.6 cm, on front of thigh.

[0045] Eight senile actinic keratosis lesions: AK 1- AK 8. Typical senile actinic keratoses that grew over two to four years. All were brown with a rough surface that did not flake off, protruded 2-3mm, and were around 12 mm diameter (but one was 2.5 x 1.2 cm), there were 5 on the back, 3 on the upper thigh.

[0046] One actinic keratosis lesion: AK 9. Cream colored actinic keratosis that formed a 4 mm horn. It grew over two years. 1.2 x 1 cm, on neck behind ear. [0047] Two nevus lesions: Nevus N 1 , slightly pigmented soft mole with a slightly roughened surface that had grown over three or more years, 9 mm diameter protruding about 6mm, on outer surface of elbow; and Nevus N2, over the angle of the jaw about 11 mm in diameter but otherwise similar to Nl .

[0048] One basal cell carcinoma lesion: BCC 1. Basal cell carcinoma that had been growing for one month before treatment started. 5 mm diameter, on forehead.

[0049] One melanotic nevus lesion: M 1. Melanotic nevus on the forehead was about 3-4 mm diameter. It had been present for several years. Two years ago it increased in size by 25% over one month during which it become blacker, the base had reddened and it was itching.

[0050] Subjects:

[0051] The lesions were found and treated within the past three years on two unrelated healthy 80 ± 3 year old subjects, one of whom was not taking any medication, the other, no medication except the lowest dose of budesonide and formoterol; both gave informed consent.

[0052] Diagnosis:

[0053] None of the diagnoses were verified histologically but there were confirmed by two independent medical doctors (MDs) as most probable and the SK 1 and SK 2 lesions by a dermatologist.

[0054] Results:

[0055] All the lesions were examined and photographed before treatment started and at weekly intervals thereafter. Of the 15 lesions, two (AK 1 and AK 5) were treated with undecylenic acid. Their response was unlike all the others, the lesions appeared burned and slightly scarred leaving the underlying lesion partly destroyed or covered. Treatment was discontinued after two weeks. All the other lesions regressed considerably as shown in the representative figures provided herewith. The rate of regression varied from 2 days to six months. Aside from these two, AK 1 and AK 5, all the keratotic lesions regressed under treatment, each cream was applied to at least two lesions. Usually some improvement was noticeable within a few days, it was definite after one week and then became increasingly obvious. After two weeks they were on average 50% improved, improvement being measured by the color (they became progressively paler) and by the texture (they became smoother to the touch) and became flatter, , and not visibly protuberant but still palpable.

[0056] Melanotic nevus, lesion M 1 , was treated with tolnaftate. It responded slowly to treatment taking 6 weeks before it had almost reverted to its original state (i.e., the state it had been in before its 25% increase in size and increase in pigmentation in a one month timeframe). By 9 weeks it had reverted completely to the state it had been in prior to its one month 25% increase. It has been symptomless and quiescent for the follow-up period (2 years).

[0057] Nevus, lesions N 1 and N2, were treated with tolnaftate. Nl responded to treatment slowly taking 4 weeks to show obvious regression and a further 6 weeks before becoming almost completely flat, becoming almost the same color of the surrounding skin and almost smooth. It has remained unchanged for two years. N2 responded to treatment similarly, being about 75% flatter after 4 weeks, when the color became almost the same as the surrounding skin, but in both lesions the footprint hardly changed.

[0058] Basal cell carcinoma, lesion BCC 1 , was papular, growing, with a pearly red periphery with a hard central lump, the margin was regular, and the center became blackened and ulcerated. After one month's application of tolnaftate (using the First Protocol, above), the lesion stopped growing and regressed slightly but looked active. Over the next four months as tolnaftate was applied (using the Second Protocol, above), the lesion regressed progressively. Over the sixth month, tolnaftate was administered every third day (using the Second Protocol, above) as a precaution during final healing. The superficial skin ulcer had healed, and the residual redness gradually became smaller and paler. The end result is a small hardly noticeable crater with no sign of activity after three months follow up.

[0059] Seborrhoeic keratosis, lesions SK l and SK 2, had been growing slowly for the past three-four years. Because they protruded and were rough to the touch, and were growing more melanotic a dermatologist was consulted, who predicted that it would eventually fall off even though it was presently firmly attached. One was treated for one month with the First Protocol, the second for three weeks with the Second Protocol. SK 1 was treated with tolnaftate using the First Protocol, after one week it was half the size and half as dark and continued to regress over 4 weeks. After this time it had the same footprint but looked less pigmented. The lesion did not protrude, nor did it feel rough to touch. SK 2 was treated with terbinafine with the Second Protocol. It regressed rapidly being noticeably reduced in height and in color within two days.

[0060] Systemic administration. It is believed that systemic administration (for example, oral, intrathecal, intramuscular, intravenous, intraperitoneal, suppository, or sublingual administration) may also be effective in treating neoplastic lesions of the epithelium. For systemic administration, the dosage should be calibrated to achieve a desired concentration in serum. The desired concentration can be determined experimentally, and a reasonable initial target concentration of the anti- fungal agent in serum is 1% by weight. The actual dosage required to achieve a target concentration will depend on a number of factors, as known in the art, including the extent of albumin binding of the agent. For assistance in making this determination, there is a known data point in that one hour after intravenous doses of 50 mg/day fluconazole for 6 days or 100 mg/day for seven days to healthy subjects, the cerebrospinal fluid concentrations of fluconazole were 1.26 mg/L and 2.74 mg/L, respectively. These values were approximately 52% and 62% those of serum. Four patients with an initial clinical diagnosis of meningitis also had significant concentrations of fluconazole in the cerebrospinal fluid. See The Journal of Clinical Pharmacology, Vol. 28, Issue 4, pages 363-366, April 1988. Of course, dosage amounts can be tested first in laboratory animals before being used on humans. See "Guidance for Industry: Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers", US Dept. of Health and Human Services, FDA, CDER, July 2005.

[0061] In addition to the types of anti-fungal agents described above, it is expected that other anti-fungal agents may find application in embodiments of the present invention. For example, echinocandins (such as anidulafungin, caspofungin, and micafungin) inhibit the synthesis of glucan in the cell wall of fungi via the enzyme 1 ,3 -beta glucan synthase.

[0062] Proposed theory: Despite their static sordid circumstances, fungi comprise their own 400 million year old taxonomic kingdom which has successfully colonized the oceans and the dry land. One cubic meter of soil may contain up to 20,000 km of mycelia which can grow at 11.6 mm per second. Fungi invented antibiotics, such as penicillin, toxins such as ergot, pesticides and carcinogens including aflatoxin, one of the most potent carcinogens known. Consequently they can disrupt the life of bacteria, plants and animals and degrade them, absorbing their constituents which they are incapable of metabolising. Genomic methodology has recently (Nature 2013, 498, 367-370) revealed that humans tend to be colonized by many fungal species with tremendous diversity. Just as whales accumulate barnacles with age, humans accumulate fungi. The mechanism of action of the anti-fungals of the present invention is unknown. However, it may be that inhibitors of fungal growth might inhibit the growth rate of keratinocytes, which are the cells from which all these neoplasms arise.

[0063] The embodiments of the invention described above are intended to be merely exemplary; numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.

Claims

What is claimed is:

1. A method of treating a neoplastic lesion in epithelium of a mammalian subject comprising administering locally to a site of the lesion a therapeutically effective amount of a composition including a non-allium anti- fungal agent.

2. A method according to claim 1, wherein the neoplastic lesion is a carcinoma.

3. A method according to claim 2, wherein the carcinoma is a basal cell carcinoma or a squamous cell carcinoma.

4. A method according to claim 1, wherein the neoplastic lesion is a melanoma.

5. A method according to claim 1, wherein the neoplastic lesion is a hyperkeratotic lesion.

6. A method according to claim 1, wherein the neoplastic lesion is a nevus.

7. A method according to claim 1, wherein the subject is a human.

8. A method according to claim 1, wherein administering includes applying the composition topically to the lesion.

9. A method according to claiml, wherein administering includes injecting the composition into the lesion.

10. A method according to claiml, wherein administering includes injecting the composition subcutaneously.

11. A method according to claim 1 , wherein the anti- fungal agent is synthetic.

12. A method according to claim 1, wherein the anti- fungal agent inhibits synthesis of ergo sterol in a fungus.

13. A method according to claim 12, wherein the anti- fungal agent inhibits squalene monoxygenase.

14. A method according to claim 12, wherein the anti-fungal agent inhibits lanosterol 14 alpha-demethylase.

15. A method according to claim 1, wherein the anti- fungal agent inhibits 1,3-beta glucan synthase.

16. A method according to claim 1, wherein the composition further comprises a chemotherapeutic agent.

17. A method according to claim 1, wherein the composition further comprises a cosmetic agent.

18. A method of treating a neoplastic lesion in epithelium of a mammalian subject comprising administering systemically a therapeutically effective amount of a composition including a non-allium anti- fungal agent, wherein the anti- fungal agent is selected from the group consisting of an agent that inhibits squalene monoxygenase and an agent that inhibits 1,3-beta glucan synthase.

19. A method according to claim 18, wherein the neoplastic lesion is a carcinoma.

20. A method according to claim 19, wherein the carcinoma is a basal cell carcinoma or a squamous cell carcinoma.

21. A method according to claim 18, wherein the neoplastic lesion is a melanoma.

22. A method according to claim 18, wherein the neoplastic lesion is a hyperkeratotic lesion.

23. A method according to claim 18, wherein the neoplastic lesion is a nevus.

24. A method according to claim 18, wherein the subject is a human.

25. A method according to claim 18, wherein administering includes injecting the composition intravenously.

26. A method according to claim 18, wherein administering includes injecting the composition intraperitoneally.

27. A method according to claim 18, wherein administering includes injecting the composition intramuscularly.

28. A method according to claim 18, wherein administering includes injecting the composition intrathecally.

29. A method according to claim 18, wherein administering includes oral administration of the composition.