Patent Grant
11478457
This invention relates to inhibitors of UDP-glucose dehydrogenase, and more particularly to UDP-glucose dehydrogenase inhibitors that are useful in the treatment of prostate cancer.
Prostate cancer is the most frequently diagnosed cancer among men in the United States, and the second most frequent cause of cancer death. One of the first line treatments for inoperable or locally advanced cancers is androgen deprivation therapy, since the cells of the prostate normally depend on circulating androgen hormones for survival. This treatment fails in approximately 20% of cases treated this way, and leads to even more aggressive cancer, termed castration resistant prostate cancer (CRPC).
The present application provides, inter alia, a method of treating prostate cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a compound provided in Table 1, or a pharmaceutically acceptable salt thereof.
In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the compound is administered in combination with one or more additional therapies. In some embodiments, at least one of the one or more additional therapies comprises administration of a chemotherapeutic agent. In some embodiments, at least one of the one or more additional therapeutic agents comprises androgen deprivation therapy. In some embodiments, the compound is administered prior to the one or more additional therapies. In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of modulating an activity of UDP-glucose dehydrogenase (UGDH) in a cell, the method comprising contacting the cell with an effective amount of a compound provided in Table 1, or a pharmaceutically acceptable salt thereof.
In some embodiments, the modulating an activity of UDP-glucose dehydrogenase (UGDH) comprises inhibiting UDP-glucose dehydrogenase (UGDH).
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of treating a prostate cancer mediated by UDP-glucose dehydrogenase (UGDH) in a patient in need thereof, the method comprising administering a therapeutically effective amount of a compound provided in Table 1, or a pharmaceutically acceptable salt thereof.
In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of predicting patient response to prostate cancer therapy, comprising:
a) obtaining a biopsy sample from the patient, wherein the biopsy sample comprises prostate cancer cells and non-cancerous tissue cells; and
b) comparing the UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and the non-cancerous tissue cells;
wherein if the UDP-glucose dehydrogenase (UGDH) expression is greater in the prostate cancer cells compared to the UDP-glucose dehydrogenase (UGDH) expression in the non-cancerous tissue cells, then the patient is more likely to respond to the prostate cancer therapy.
In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the comparing comprises determining the ratio of UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and UDP-glucose dehydrogenase (UGDH) expression in the non-cancerous tissue cells.
In some embodiments, the prostate cancer therapy comprises androgen deprivation therapy. In some embodiments, the prostate cancer therapy is androgen deprivation therapy.
The present application further provides a method of treating a prostate cancer mediated by UDP-glucose dehydrogenase (UGDH) in a patient in need thereof, the method comprising:
a) obtaining a biopsy sample from the patient, wherein the biopsy sample comprises prostate cancer cells and non-cancerous tissue cells;
b) comparing the UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and the non-cancerous tissue cells; and
c) if the prostate cancer is determined to be associated with one or more of overexpression and amplification of UDP-glucose dehydrogenase (UGDH) in the prostate cancer cells, administering a therapeutically effective amount of a compound provided in Table 1, or a pharmaceutically acceptable salt thereof.
In some embodiments, the comparing comprises determining the ratio of UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and UDP-glucose dehydrogenase (UGDH) expression in the non-cancerous tissue cells.
In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the method further comprises administration of one or more additional therapies. In some embodiments, at least one of the one or more additional therapies comprises administration of a chemotherapeutic agent. In some embodiments, at least one of the one or more additional therapeutic agents comprises administration of androgen deprivation therapy.
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of improving the efficacy of androgen deprivation therapy in a patient, comprising administering to the patient a therapeutically effective amount of a UDP-glucose dehydrogenase (UGDH) inhibitor.
In some embodiments, the androgen deprivation therapy is administered for the treatment of prostate cancer in the patient. In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the UDP-glucose dehydrogenase (UGDH) inhibitor is selected from a compound provided in Table 1, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.
UDP-glucose dehydrogenase (UGDH) catalyzes the NAD+-dependent, two-step oxidation of UDP-glucose to UDP-glucuronic acid, an essential precursor for hyaluronan synthesis by HAS enzymes, other glycosaminoglycan/proteoglycan production in the Golgi, and glucuronidation of steroid hormones by UGTs for solubilization and excretion (see, e.g., Prydz et al., J. Cell Sci. 2000, 113, 193-205; Fraser et al., J. Intern. Med. 1997, 242, 27-33; Guillemette C., Pharmacogenomics J. 2003, 3, 136-158; and King et al., Toxicol. Sci. 2001, 61, 49-53). High levels of UGDH expression are specific to the liver and prostate in males, and prostate tumor progression has been correlated with a loss of UGDH regulation. Accordingly, the present application provides inhibitors of UGDH that are useful in the treatment of prostate cancer and methods for predicting the efficacy of androgen deprivation therapy.
For the terms “for example” and “such as” and grammatical equivalences thereof, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. As used herein, the term “about” is meant to account for variations due to experimental error. All measurements reported herein are understood to be modified by the term “about”, whether or not the term is explicitly used, unless explicitly stated otherwise. As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise.
A “therapeutically effective amount” of a conjugate with respect to the subject method of treatment, refers to an amount of the conjugate(s) in a preparation which, when administered as part of a desired dosage regimen (to a patient, e.g., a human) alleviates a symptom, ameliorates a condition, or slows the onset of disease conditions according to clinically acceptable standards for the disorder or condition to be treated or the cosmetic purpose, e.g., at a reasonable benefit/risk ratio applicable to any medical treatment.
As used herein, the term “treating” or “treatment” includes reversing, reducing, or arresting the symptoms, clinical signs, and underlying pathology of a condition in manner to improve or stabilize a patient's condition.
Compounds and Pharmaceutical Compositions
The present application provides, inter alia, compounds that are useful as UDP-glucose dehydrogenase (UGDH) inhibitors. In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1) (i.e., compound (1) or inhibitor (1)) is also referred to as inhibitor 5210344. In some embodiments, the compound 3-[5-(2-thienyl)-2-furyl]propanoic acid (31) (i.e. compound (31) or inhibitor (31)) is also referred to as inhibitor 6847944.
In some embodiments, a compound is selected from the group provided in Table 1.
The present application further provides a pharmaceutical composition comprising a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.
When employed as pharmaceuticals, the compounds provided herein can be administered in the form of pharmaceutical compositions; thus, the methods described herein can include administering pharmaceutical compositions provided herein.
These compositions can be prepared as described herein or elsewhere, and can be administered by a variety of routes, depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration may include, but is not limited to intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection or infusion; or intracranial, (e.g., intrathecal, intraocular, or intraventricular) administration. Parenteral administration can be in the form of a single bolus dose, or may be, for example, by a continuous perfusion pump. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
In making the compositions provided herein, the active ingredient is typically mixed with an excipient, diluted by an excipient or enclosed within such a carrier in the form of, for example, a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.
Some examples of suitable excipients include, without limitation, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The formulations can additionally include, without limitation, lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl- and propylhydroxy-benzoates; sweetening agents; flavoring agents, or combinations thereof.
The active compounds can be effective over a wide dosage range and are generally administered in a pharmaceutically effective amount. It will be understood, however, that the amount of the compound actually administered and the schedule of administration will usually be determined by a physician, according to the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual subject, the severity of the subject's symptoms, and the like.
Methods of Use and Combination Therapies
The present application further provides methods of treating prostate cancer in a patient in need thereof. As used herein, the term “patient” refers to any animal, including mammals, for example, mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, primates, and humans. In some embodiments, the patient is a human. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
In some embodiments, the prostate cancer comprises a cancer selected from the group consisting of acinar adenocarcinoma, atropic adenocarcinoma, foamy adenocarcinoma, colloid adenocarcinoma, signet ring carcinoma, ductal adenocarcinoma transitional cell (or urothelial) cancer, squamous cell cancer, carcinoid, small cell cancer, sarcoma cancer, sarcomatoid cancer, and castration resistant prostate cancer (CRPC). In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof is administered in combination with one or more additional therapies. In some embodiments, at least one of the one or more additional therapies is selected from the group consisting of administration of a chemotherapeutic agent, radiation therapy, a surgical procedure, androgen deprivation therapy, or any combination thereof. In some embodiments, at least one of the one or more additional therapies comprises administration of at least one chemotherapeutic agent. In some embodiments, at least one of the one or more additional therapies comprises androgen deprivation therapy. In some embodiments, at least one of the one or more additional therapies is androgen deprivation therapy. In some embodiments, a compound provided herein, or a pharmaceutically acceptable salt thereof, is administered prior to the one or more additional therapies. In some embodiments, a compound provided herein, or pharmaceutically acceptable salt thereof, is administered concurrently with the one or more additional therapies. In some embodiments, a compound provided herein, or a pharmaceutically acceptable salt thereof, is administered after the one or more additional therapies.
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof, and at least one of the one or more additional therapies is androgen deprivation therapy. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof, and at least one of the one or more additional therapies is androgen deprivation therapy.
In some embodiments, the method comprises:
i) administering to the patient a therapeutically effective amount of 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof; and
ii) administering androgen deprivation therapy.
In some embodiments, the method comprises:
i) administering to the patient a therapeutically effective amount of 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof; and
ii) administering androgen deprivation therapy.
The present application further provides a method of modulating an activity of UDP-glucose dehydrogenase (UGDH) in a cell, the method comprising contacting the cell with an effective amount of a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the modulating an activity of UDP-glucose dehydrogenase (UGDH) comprises inhibiting UDP-glucose dehydrogenase (UGDH). In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of treating a prostate cancer mediated by UDP-glucose dehydrogenase (UGDH) in a patient in need thereof, the method comprising administering a therapeutically effective amount of a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof.
In some embodiments, the prostate cancer comprises a cancer selected from the group consisting of acinar adenocarcinoma, atropic adenocarcinoma, foamy adenocarcinoma, colloid adenocarcinoma, signet ring carcinoma, ductal adenocarcinoma transitional cell (or urothelial) cancer, squamous cell cancer, carcinoid, small cell cancer, sarcoma cancer, sarcomatoid cancer, and castration resistant prostate cancer (CRPC). In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of predicting patient response to prostate cancer therapy, comprising:
a) obtaining a biopsy sample from the patient, wherein the biopsy sample comprises prostate cancer cells and non-cancerous tissue cells; and
b) comparing the UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and the non-cancerous tissue cells;
wherein when the UDP-glucose dehydrogenase (UGDH) expression is greater in the prostate cancer cells compared to the UDP-glucose dehydrogenase (UGDH) expression in the non-cancerous tissue cells, then the patient is more likely to respond to the prostate cancer therapy.
In some embodiments, the prostate cancer cells comprise a prostate cancer selected from the group consisting of acinar adenocarcinoma cells, atropic adenocarcinoma cells, foamy adenocarcinoma cells, colloid adenocarcinoma cells, signet ring carcinoma cells, ductal adenocarcinoma transitional cell (or urothelial) cancer cells, squamous cell cancer cells, carcinoid cells, small cell cancer cells, sarcoma cancer cells, sarcomatoid cancer cells, and castration resistant prostate cancer (CRPC) cells. In some embodiments, the prostate cancer cells comprise castration resistant prostate cancer (CRPC) cells. In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
As used herein, the term “non-cancerous tissue cells” refers to non-cancerous tissue cells in the area surrounding the prostate cancer cells. For example, non-cancerous tissue cells may refer to non-cancerous prostate tissue cells, non-cancerous acini, and normal-appearing acini (NAA). In some embodiments, the non-cancerous or normal-appearing acini are selected from the group consisting of acini of the stomach, acini of the sebaceous gland of the scalp, acini of the liver, acini of the lung, acini of the lacrimal gland, acini of mammary gland, acini of the pancreas, and acini of the prostate. In some embodiments, the non-cancerous tissue cells comprise prostate tissue cells. In some embodiments, the non-cancerous tissue cells are prostate tissue cells. In some embodiments, the non-cancerous tissue cells comprise non-cancerous acini or normal-appearing acini (NAA). In some embodiments, the non-cancerous cells comprise normal-appearing acini. Examples of normal-appearing acini that may be used in the method provided herein may be found, for example, in Huang et al., Int. J. Cancer, 2010, 126(5), 315-327, the disclosure of which is incorporated herein in its entirety.
In some embodiments, the comparing comprises determining the ratio of UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and UDP-glucose dehydrogenase (UGDH) expression in the non-cancerous tissue cells. In some embodiments, the comparing comprises:
a) immunofluorescence staining of the biopsy sample; and
b) quantifying the fluorescence pixel intensity of acini within the prostate cancer cells and the non-cancerous tissue cells.
In some embodiments, the quantifying comprises determining the average mean pixel intensity acini within the prostate cancer cells and acini within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)).
In some embodiments, the patient is more likely to respond to prostate cancer therapy when the average mean pixel intensity of acini within the prostate cancer cells is at least about 10% greater than the average mean pixel intensity of acini within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)), for example, at least about 10%, at least about 15% at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%. In some embodiments, the patient is more likely to respond to prostate cancer therapy when the average mean pixel intensity of acini within the prostate cancer cells is at least about 15% greater than the average mean pixel intensity of acini within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)).
In some embodiments, the prostate cancer therapy is selected from the group consisting of administration of a chemotherapeutic agent, radiation therapy, a surgical procedure, androgen deprivation therapy, or any combination thereof. In some embodiments, a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof, is administered prior to the prostate cancer therapy. In some embodiments, a compound provided herein, or pharmaceutically acceptable salt thereof, is administered concurrently with the prostate cancer therapy. In some embodiments, a compound provided herein, or a pharmaceutically acceptable salt thereof, is administered after the prostate cancer therapy. In some embodiments, the prostate cancer therapy comprises administration of at least one chemotherapeutic agent. In some embodiments, the prostate cancer therapy comprises androgen deprivation therapy. In some embodiments, the prostate cancer therapy is androgen deprivation therapy.
The present application further provides a method of treating a prostate cancer mediated by UDP-glucose dehydrogenase (UGDH) in a patient in need thereof, the method comprising:
a) obtaining a biopsy sample from the patient, wherein the biopsy sample comprises prostate cancer cells and non-cancerous tissue cells;
b) comparing the UDP-glucose dehydrogenase (UGDH) expression in the prostate cancer cells and the non-cancerous tissue cells; and
c) if the prostate cancer is determined to be associated with one or more of overexpression and amplification of UDP-glucose dehydrogenase (UGDH) in the prostate cancer cells compared to the non-cancerous tissue cells, administering a therapeutically effective amount of a compound provided herein, or a pharmaceutically acceptable salt thereof.
In some embodiments, the prostate cancer is determined to be associated with one or more of overexpression and amplification of UDP-glucose dehydrogenase (UGDH) in the prostate cancer cells compared to the non-cancerous tissue cells when the average mean pixel intensity of acini within the prostate cancer cells is at least about 10% greater than the average mean pixel intensity of acini within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)), for example, at least about 10%, at least about 15% at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%. In some embodiments, the patient is more likely to respond to prostate cancer therapy when the average mean pixel intensity of acini within the prostate cancer cells is at least about 15% greater than the average mean pixel intensity of acini within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)).
In some embodiments, the prostate cancer is determined to be associated with one or more of overexpression and amplification of UDP-glucose dehydrogenase (UGDH) in the prostate cancer cells compared to the non-cancerous tissue cells when the expression of UGDH within the prostate cancer cells is at least about 10% greater than the expression of UGDH within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)), for example, at least about 10%, at least about 15% at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100%. In some embodiments, the patient is more likely to respond to prostate cancer therapy when the expression of UGDH is within the prostate cancer cells is at least about 15% greater than the expression of UGDH within the non-cancerous tissue cells (e.g., non-cancerous acini or normal-appearing acini (NAA)).
In some embodiments, the method further comprises administration of one or more additional therapies. In some embodiments, at least one of the one or more additional therapies is selected from the group consisting of administration of a chemotherapeutic agent, radiation therapy, a surgical procedure, androgen deprivation therapy, or any combination thereof. In some embodiments, at least one of the one or more additional therapies administering at least one chemotherapeutic agent. In some embodiments, at least one of the one or more additional therapies comprises androgen deprivation therapy. In some embodiments, at least one of the one or more additional therapies is androgen deprivation therapy. In some embodiments, a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof, is administered prior to the one or more additional therapies. In some embodiments, a compound provided herein, or pharmaceutically acceptable salt thereof, is administered concurrently with the one or more additional therapies. In some embodiments, a compound provided herein, or a pharmaceutically acceptable salt thereof, is administered after the one or more additional therapies.
In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The present application further provides a method of improving the efficacy of androgen deprivation therapy in a patient, comprising administering to the patient a therapeutically effective amount of a UDP-glucose dehydrogenase (UGDH) inhibitor.
In some embodiments, the prostate cancer comprises a cancer selected from the group consisting of acinar adenocarcinoma, atropic adenocarcinoma, foamy adenocarcinoma, colloid adenocarcinoma, signet ring carcinoma, ductal adenocarcinoma transitional cell (or urothelial) cancer, squamous cell cancer, carcinoid, small cell cancer, sarcoma cancer, sarcomatoid cancer, and castration resistant prostate cancer (CRPC). In some embodiments, the prostate cancer is castration resistant prostate cancer (CRPC).
In some embodiments, the UDP-glucose dehydrogenase (UGDH) inhibitor is selected from a compound provided herein (e.g., a compound provided in Table 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 2,2′-[1-(4-amino-1,2,5-oxadiazol-3-yl)-1H-1,2,3-triazole-4,5-diyl]di(2-butanol) (1), or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is 3-[5-(2-thienyl)-2-furyl]propanoic acid (31), or a pharmaceutically acceptable salt thereof.
The invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of non-critical parameters which can be changed or modified to yield essentially the same results.
i. Reagents
ii. Cocktails
Enzyme Buffer 2.8×
14 μg/mL enzyme;
0.56% BSA; and
In PBS, pH 7.4.
Preparation of 1 liter: Prepare enzyme in polypropylene. Do not let enzyme touch glass or polystyrene. Because the enzyme is the least stable of all reagents used in the assay, it should be prepared last. Enzyme buffer should be kept cold if any time passes before use.
700 μM NAD+;
92.4 μM UDP-glucose; and
In PBS, pH 7.4.
Preparation of 1 L:
70 μM UDP-xylose;
2.5% DMSO; and
In PBS, pH 7.4
Preparation of 100 mL:
2.5% DMSO; and
In PBS, pH 7.4.
Preparation of 100 mL:
1. Preparation of the Compound-Plates: Add Positive & Negative Controls
2. First Addition: Add Enzyme to Compounds
3. First Read: Measure Baseline (Compound) Fluorescence.
4. Second Addition: Add Substrates to Enzyme & Compounds
5. Second Read: Measure NADH Levels
6. Standard Curves:
Plates can be run in batches up to 25.
iv. Data-Analysis
All data is converted to NADH concentration using the standard curve generated each day. All data is converted to: [second read−the first read]. Because NADH is generated in the reaction, an increase in NADH signals enzyme activity. Percent of Controls (POC) expresses the activity of a compound relative to the positive and negative controls (Equation 1).
Note that the positive control wells (with UDP-xylose) will have lower value (less NADH) than will negative control wells. So the denominator in this equation will be a negative number. An active (inhibitory) compound will also result in a lower value, so the numerator will also be negative. Thus, the higher the POC, the more inhibitory the compound is (see e.g., Table 2).
Table 3 shows POC values for compounds screened in the high throughput assay.
Kinetic parameters of WT and mutant UGDH are shown below in Table 4. The Km and Vmax for the substrate (UDP-glucose) and cofactor (NAD+) was determined by a nonlinear regression fit of initial velocity vs. substrate/cofactor concentration. T325A and T325D are engineered inducible hexameric and obligate dimeric UGDH species, respectively.
Table 5 shows inhibition data for UDP-xylose, inhibitor (1) (i.e., 5210344), and inhibitor (31) (i.e., 6847944). IC50 values for UDP-xylose and inhibitor (1) were determined using Km concentrations of UDP-glucose and NAD+ (50 μM UDP-glc and 1 mM NAD+). Ki values were determined by varying [UDP-glc] and holding NAD+ at saturating concentrations. UDP-xylose is a more potent inhibitor than the other compounds. Inhibitor (1) may have greatest effect on the dimer. Inhibitor (31) appears to require the ability for hexamer formation to inhibit which may suggest interference at the dimer-dimer interface as mechanism of action.
UDP-Xylose Enhances Thermal Stability
Inhibitor (1) Selectively Affects Thermal Stability
Inhibitor (31) has Negligible Effect on Thermal Stability
UDP-xylose stabilizes UGDH T325A and T325D mutants against limited trypsin proteolysis, as shown in
It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.
This application claims the benefit of U.S. Provisional Application Ser. No. 62/151,869, filed Apr. 23, 2015, the disclosure of which is incorporated herein by reference in its entirety.
This invention was made with Government support under Grant Nos. 2012-31100-06031, 2013-31100-06031, 2014-31100-06031, and 2015-31100-06031 awarded by the United States Department of Agriculture and the National Institute of Food and Agriculture. The Government has certain rights in the invention.
| Number | Name | Date | Kind |
|---|---|---|---|
| 8106037 | Rubin et al. | Jan 2012 | B2 |
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