Methods for treating pancreatic cancer

Abstract

The present invention provides a method for treating pancreatic cancer. The method comprises administering an effective amount of dapanstril to a subject in need thereof. The preferred route of administration of dapanstril is oral administration. The present invention also provides a combination therapy for pancreatic cancer by administering dapanstril and gemcitabine to a patient in need thereof.

Description

[Technical field]

[0001] The present invention relates to a method for treating pancreatic cancer by administering to a subject in need thereof an effective amount of dapansutrile. [Background technology]

[0002] Pancreatic ductal adenocarcinoma (PDAC) accounts for 90% of pancreatic cancer cases and is the third leading cause of cancer deaths in the United States and the seventh leading cause worldwide (PMID:35020204, 30834048). With a reported increase in incidence over the years, pancreatic cancer is predicted to become the second leading cause of cancer-related deaths in the United States by 2030 (PMID:34547082). The lack of effective screening methods and non-specific symptoms even at advanced stages are major limitations in the management of this disease that contribute to the extremely grim prognosis of pancreatic cancer patients (5-year survival rate reached 11% for the first time in 2022). To date, at the time of diagnosis, only 10-15% of patients present with localized disease and are candidates for surgery, while the remaining 85-90% of patients have metastatic disease (stage IV) or locally advanced disease (stages II and III) that is not suitable for resection (PMID:34547082). Treatment options for patients with unresectable disease are very limited, including radiation therapy and chemotherapy regimens including gemcitabine, nab-paclitaxel, and FOLFIRINOX. Given that standard therapies have only a minimal impact on overall survival for cancer patients with advanced disease and that serious adverse events associated with these treatments are common, novel treatment options represent a medical urgency for these patients.

[0003] Inflammation has been shown to affect PDAC growth, response to therapy, and metastasis, and several inflammatory cytokines have been found to be increased in PDAC patients (PMID:25897428). Among these, the inflammatory cytokine IL-1β has been shown to be upregulated in PDAC, promote desmoplasia, immunosuppression, and correlate with patient responsiveness to chemotherapy (PMID:23591198;31915130;26500238). IL-1β is initially synthesized as a biologically inactive proform that requires processing (ref). IL-1β maturation is mostly controlled by conserved cytoplasmic pattern recognition receptors (PRRs) of the nucleotide-binding and oligomerization domain NOD-like receptor (NLR) family. In the pancreatic context, one NLRs member, NLRP3, has been shown to mediate inflammation in acute pancreatitis, which is recognized as a high-risk factor for the development of PDAC (PMID:33228173). Furthermore, NLRP3 signaling has been shown to be involved in tumor progression in mouse models of PDAC (PMID:28442553).

[0004] There is a need for methods for treating pancreatic cancer that are effective and desirably free of significant side effects. [Brief description of the drawings]

[0005] [Figure 1] Figure 1 shows that pancreatic cancer-bearing mice fed the dapanstril (OLT) diet showed reduced tumor volume (****=p<0.0001) and reduced tumor weight (**=p<0.01) compared to mice fed the standard (STD) diet (Figure 1). [Diagram 2] Figure 2 shows that pancreatic cancer-bearing mice treated with Dapanstril (OLT) + Gemcitabine (GEM) had significantly further reduced tumor volume and tumor weight compared to pancreatic cancer-bearing mice treated with OLT alone or GEM alone. ****=p<0.0001, ***=p<0.001, **=p<0.01, *=p<0.05. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0006] Detailed Description of the Invention The present invention relates to a method for treating pancreatic cancer, such as pancreatic ductal adenocarcinoma, using dapanstril, an NLRP3 inhibitor.

[0007] NLRP3 (NOD-like receptor family, pyrin domain containing 3), also known as NALP3 or cryopyrin, is one of the sensors of the inflammasome, a macromolecular structure involved in the processing of interleukin-1β (IL-1β) and IL-18. Activation of NLRP3 triggers the recruitment of ASC (apoptosis-associated speck-like protein containing carboxy-terminal caspase recruitment domain) and caspase-1, leading to the formation of the inflammasome and eventual cell death.

[0008] NLRP3 is highly expressed in pancreatic cancer tissues compared to normal pancreas. NLRP3 is a cytoplasmic receptor that determines the maturation of biologically inactive inflammatory cytokines IL-1β and IL-18 into their biologically active forms after activation. Once activated, NLRP3 forms inflammasomes that mediate the processing of pro-IL-1β and pro-IL-18 into their biologically active forms. We found that NLRP3 inflammasomes are formed in human PDAC samples. Consistently, IL-1β levels are elevated in PDAC samples.

[0009] compound The present invention uses the purified compound of dapanstril (3-methanesulfonyl-propionitrile), or a pharma- ceutically acceptable solvate thereof, to treat pancreatic cancer. [ka] Dapanstril

[0010] Dapanstril is a small, synthetic molecule of β-sulfonylnitrile that has been shown to selectively inhibit the NLRP3 inflammasome and is safe when administered orally to healthy individuals.

[0011] As used herein, "solvates" are addition complexes in which a compound is combined in a fixed ratio with an acceptable co-solvent, including, but not limited to, water, acetic acid, ethanol, and other suitable organic solvents.

[0012] Pharmaceutical Compositions Generally, the active compound dapanstril, or a pharma- ceutically acceptable solvate thereof, in the pharmaceutical composition is in an amount of about 0.1-5% for injectable formulations, about 1-90% for tablet formulations, about 1-100% for capsule formulations, about 0.01-20%, 0.05-20%, 0.1-20%, 0.2-15%, 0.5-10%, or 1-5% (w / w) for topical formulations, and about 0.1-5% for patch formulations.

[0013] As used in this application, "about" refers to ±10% of the stated value.

[0014] The pharma- ceutically acceptable carrier, which is an inactive ingredient, can be selected by those skilled in the art according to conventional standards. Pharmaceutically acceptable carriers include, but are not limited to, non-aqueous-based solutions, suspensions, emulsions, microemulsions, micellar solutions, gels, and ointments. Pharmaceutically acceptable carriers also include physiological saline and aqueous electrolyte solutions; ionic and non-ionic osmotic agents, such as sodium chloride, potassium chloride, glycerol, and dextrose; pH adjusters and buffers, such as hydroxide salts, phosphate salts, citrate salts, acetate salts, borates, and trolamine; antioxidants, such as bisulfite, sulfite, metabisulfite, and thiosulfite salts, acids and / or bases, ascorbic acid, acetylcysteine, cysteine, glutathione, butylated hydroxyanisole, butylated hydroxytoluene, tocopherol, and ascorbyl palmitate; surfactants, such as lecithin, acetylcysteine, cysteine, glutathione, butylated hydroxyanisole, butylated hydroxytoluene, tocopherol, and ascorbyl palmitate; and surfactants, such as lecithin, acetylcysteine ​​... phospholipids, including phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol; poloxamers and proxamines; polysorbates, such as polysorbate 80, polysorbate 60, and polysorbate 20; polyethers, such as polyethylene glycol and polypropylene glycol; polyvinyls, such as polyvinyl alcohol and povidone; cellulose derivatives, such as methylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose, and hydroxypropylmethylcellulose, and salts thereof; petroleum derivatives, such as cellulose acetate, cellulose acetate, cellulose esters ... The cosmetic composition may also include ingredients such as, but not limited to, fatty acids, oleic acid derivatives, such as mineral oil and white petroleum derivatives; fats and oils, such as lanolin, peanut oil, palm oil, soybean oil, and the like; mono-, di-, and triglycerides; polymers of acrylic acid, such as carboxypolymethylene gel, and hydrophobically modified cross-linked acrylate copolymer, and the like; polysaccharides, such as dextran, and glycosaminoglycans, such as sodium hyaluronate, and the like.Such pharma- ceutically acceptable carriers can be preserved against bacterial contamination using well-known preservatives, including, but not limited to, benzalkonium chloride, ethylenediaminetetraacetic acid and its salts, benzethonium chloride, chlorhexidine, chlorobutanol, methylparaben, thimerosal, and phenylethyl alcohol, or can be formulated as unpreserved formulations for either single or multiple use.

[0015] For example, tablet or capsule formulations of dapanstril may contain other excipients that are not biologically active and do not react with the active compound. Tablet excipients include fillers, binders, lubricants and glidants, disintegrants, wetting agents, and release rate modifiers. Binders promote the adhesion of the particles of the formulation and are important for tablet formulations. Examples of binders include, but are not limited to, carboxymethylcellulose, cellulose, ethylcellulose, hydroxypropylmethylcellulose, methylcellulose, karaya gum, starch, and tragacanth gum, poly(acrylic acid), and polyvinylpyrrolidone.

[0016] For example, a patch formulation of dapanstril may contain several inactive ingredients, such as 1,3-butylene glycol, dihydroxyaluminum aminoacetate, disodium edetate, D-sorbitol, gelatin, kaolin, methylparaben, polysorbate 80, povidone, propylene glycol, propylparaben, sodium carboxymethylcellulose, sodium polyacrylate, tartaric acid, titanium dioxide, and purified water, etc. The patch formulation may also contain a skin permeation enhancer, such as a lactate ester (e.g., lauryl lactate) or diethylene glycol monoethyl ether.

[0017] Topical formulations containing dapanstril can be in the form of gel, cream, lotion, liquid, emulsion, ointment, spray, solution, suspension.Inactive ingredients in topical formulations include, but are not limited to, for example, lauryl lactate (emollient / penetration enhancer), diethylene glycol monoethyl ether (emollient / penetration enhancer), DMSO (solubility enhancer), silicone elastomer (rheology / texture modifier), caprylic / capric triglyceride (emollient), octisalate (emollient / UV filter), silicone fluid (emollient / diluent), squalene (emollient), sunflower oil (emollient), and silicon dioxide (thickener).In one embodiment, diethylene glycol monoethyl ether is included in topical gel formulations.

[0018] How to use By inhibiting the assembly of the NLRP3 inflammasome, dapanstril prevents the production and / or release of the pro-inflammatory cytokine IL-1β, ultimately reducing the growth of pancreatic tumors.

[0019] The present invention is directed to a method for treating pancreatic cancer, particularly PDAC.The method comprises administering an effective amount of dapanstril to a subject in need thereof.As used herein, "effective amount" refers to an amount that is effective for treating disease by improving pathology and / or reducing, improving, and / or eliminating symptoms of disease.For example, an effective amount is an amount that reduces the proliferation of pancreatic cancer (reducing tumor size and / or reducing tumor weight).

[0020] In one embodiment, a patient suffering from pancreatic cancer is administered dapanstril without receiving immune cells, such as cytokine-induced killer cells.

[0021] The pharmaceutical composition containing dapanstril can be applied by systemic administration or local administration. Systemic administration includes, but is not limited to, oral administration, parenteral administration (e.g., intravenous administration, intramuscular administration, subcutaneous administration, or rectal administration, etc.), and inhalation administration. In systemic administration, the active compound first reaches the plasma and then distributes to the target tissue. Oral administration is the preferred administration route of the present invention. Local administration includes topical administration.

[0022] The dosage of the dapanstril composition may vary based on the extent of the tumor in question and the individual response of each patient. With systemic administration, the plasma concentration of the delivered active compound may vary; generally, it is within the range of 1×10 -10 ~1×10 -4 moles / liter, preferably 1 x 10 -8 ~1×10 -5 It is in moles per liter.

[0023] In one embodiment, the pharmaceutical composition is orally administered to a subject. The dosage for oral administration is generally at least 0.1 mg of drug / kg of subject / day, and less than 100 mg / kg / day or less than 200 mg / kg / day. For example, the dosage for oral administration is 1-100, or 5-50, or 10-50 mg / kg / day to a subject. For example, the dosage for oral administration is 100-10,000 mg / day to a subject, and preferably 500-2000, 500-4000, 500-4000, 1000-5000, 2000-5000, 2000-6000, or 2000-8000 mg / day to a subject. Dapanstril can be taken orally once, twice, three times, or four times a day, depending on the age and condition of the patient.

[0024] In one embodiment, the pharmaceutical composition is administered to a subject intravenously. The dosage for intravenous bolus injection or intravenous infusion is generally 0.03-5 or 0.03-1 mg / kg / day.

[0025] In one embodiment, the pharmaceutical composition is administered subcutaneously to a subject. The dose for subcutaneous administration is generally 0.3 to 20, 0.3 to 3, or 0.1 to 1 mg / kg / day.

[0026] In one embodiment, the composition is applied topically. The composition is applied topically at least 1-2 times a day, or 3-4 times a day, depending on the medical problem and the symptoms of the disease. Generally, the topical composition contains about 0.01-20%, or 0.05-20%, or 0.1-20%, or 0.2-15%, 0.5-10%, or 1-5% (w / w) of the active compound. Typically, 0.2-10 mL of the topical composition is applied to an individual at each time.

[0027] Those of skill in the art will recognize that a wide variety of delivery mechanisms are also suitable for the present invention.

[0028] Combination therapy Given the limited efficacy of current therapies for PDAC, finding more effective ways to treat human pancreatic cancer is an urgent need.

[0029] The present invention is further directed to a method for treating pancreatic cancer by administering dapanstril and a chemotherapeutic agent. The method comprises administering to a subject in need thereof an effective amount of dapanstril and an effective amount of a chemotherapeutic agent. Suitable chemotherapeutic agents for use with dapanstril to treat pancreatic cancer include, for example, gemcitabine, FOLFIRINOX and nab-paclitaxel.

[0030] FOLFIRINOX is a potent chemotherapy drug that includes leucovorin calcium (folinic acid), fluorouracil, irinotecan hydrochloride, and oxaliplatin.

[0031] Nab-paclitaxel, also known as Abraxane, is a chemotherapy drug that binds the chemotherapy drug paclitaxel to a protein called albumin.

[0032] Gemcitabine hydrochloride is an intravenously administered chemotherapy antimetabolite and a potent and specific oncolytic used in the treatment of cancer. The mechanism of antiproliferative activity of gemcitabine includes inhibition of DNA replication and repair by inhibiting DNA synthesis and inhibiting repair mechanisms by masked strand termination. Gemcitabine inhibits the proliferation of highly proliferative cells such as tumor cells.

[0033] The inventors have demonstrated that the combination of dapanstril and gemcitabine significantly inhibits tumor progression compared to monotherapy in animals. Gemcitabine is a chemotherapy that inhibits DNA synthesis, stopping the increase of hyperproliferative cells. Dapanstril inhibits NLRP3, enhances T cell activity, prevents inflammatory events mediated by PDAC, and ultimately reduces immunosuppression. Mechanistically, treatment with dapanstril does not affect the proliferation of non-immune cells, but significantly reduces COX2 / PGE2, a signal of a potent immunosuppressive pathway in PDAC. The independent targets of gemcitabine and dapanstril provide the basis for the beneficial effect of combination therapy. The combination of these two drugs increases the efficacy of each monotherapy because they combine two different mechanisms of inhibiting tumor cell proliferation. Combination therapy of dapanstril and gemcitabine may reduce the dose of gemcitabine and provide a long-term tolerable treatment for pancreatic cancer patients.

[0034] In one embodiment of the combination therapy, dapanstril is administered orally daily and the chemotherapy agents are administered according to their standard routes and standard dosages, or at lower dosages. For example, gemcitabine is administered at 500-1000 mg / m 2 Surface area or 800-1000mg / m 2 It is administered intravenously weekly in surface area doses.

[0035] The invention is useful for treating mammalian subjects, such as humans, horses, dogs, and cats, etc. The invention is particularly useful for treating humans.

[0036] The following examples further illustrate the invention. These examples are intended merely to be illustrative of the invention and are not to be construed as limiting. EXAMPLES

[0037] Example 1. Dapanstril reduced pancreatic cancer growth in mice. C57BL / 6 male mice (8 weeks old) were purchased from Jackson Laboratories (Bar Harbor, ME). After anesthetizing the mice and opening the abdominal cavity, the tip of the pancreatic tail was gently grasped and the pancreas / spleen was externalized laterally to expose the pancreatic body and the entire spleen. One hundred thousand KPC cells, a mouse PDAC cell line, were mixed 1:1 with Matrigel (VF=20 μl) and injected into the pancreatic tail. The pancreas was then gently internalized with blunt forceps and abdominal muscle layer, and the skin was closed separately with absorbable sutures.

[0038] After the first injection of KPC cells, tumor-bearing mice were monitored daily until sacrifice 21 days later, at which time tumor volumes and tumor weights were assessed in vehicle-treated (n=16) and OLT1177® (dapanstril)-treated (n=16) mice.

[0039] From the day of surgery onwards, mice received ad libitum oral administration of dapanstril via food pellets. The composition of the food was standard mouse chow enriched with 7.5 gr dapanstril per kg of food. The dapanstril concentration in the food was based on the observation that mice fed this chow reached mean dapanstril plasma concentrations on the order of safe levels achieved in humans in a phase I clinical trial (PMID:30075804).

[0040] At the time of sacrifice, the tumor volumes of the different groups were calculated using the modified ellipsoidal formula V = (LW 2) / 2, where L is the length of the longest tumor dimension parallel to the skin that includes the tumor midpoint, W is the length of the tumor dimension perpendicular to L and parallel to the skin, and V is in cubic millimeters (mm 3 ) tumor volume.

[0041] As a result, mice with pancreatic cancer fed the dapanstril diet showed a reduction in tumor volume ( **** =p<0.0001) and reduction in tumor weight ( ** =p<0.01) was shown (Figure 1).

[0042] Example 2. Combination therapy of dapanstril and gemcitabine Tumor-bearing mice were prepared according to Example 1 and monitored daily until sacrifice after 21 days.

[0043] Four groups of tumor-bearing mice were treated with PBS (n=8), gemcitabine (n=11), dapanstril (n=8), and dapanstril + gemcitabine (n=9). Mice treated with dapanstril were given dapanstril according to the same protocol as described in Example 1. All other three groups of mice were fed standard chow as described in Example 1.

[0044] Gemcitabine (Hospira Inc., Lake Forest, Ill.) was administered intraperitoneally (ip) at 100 mg / kg every 3 days starting 10 days after surgery. Control mice received a PBS solution.

[0045] At the time of sacrifice, tumor volume and tumor weight were assessed in the four groups of mice.

[0046] The results are shown in Figure 2. As a result, administration of gemcitabine alone and dapanstril alone significantly reduced tumor volume and tumor weight. Mice treated with gemcitabine + dapanstril showed a significant reduction in tumor volume and tumor weight compared to vehicle-treated mice and each monotherapy (gemcitabine and dapanstril). The data show that the addition of NLRP3 inhibition with dapanstril to standard treatment gemcitabine enhances the efficacy of gemcitabine alone.

[0047] reference 1. Y. Guo, et al. Cancer Res 77, 6429-6441 (2017). 2. S.Shalapour,et al.J Clin Invest 125,3347-3355(2015). 3.CADinarello.Blood 117,3720-3732(2011). 4. RNApte, et al. Cancer Metastasis Rev 25, 387-408 (2006). 5. CA Dinarello.Cancer Metastasis Rev 29,317-329(2010). 6. B.Guo,et al.Sci Rep 6,36107(2016). 7. C.Marchetti,et al.Proc Natl Acad Sci USA 115,E1530-E1539(2018).

Claims

1. A pharmaceutical composition for treating pancreatic cancer in a subject, comprising an effective amount of dapanstril, or a pharmaceutically acceptable solvate thereof.

2. The pharmaceutical composition of claim 1 , wherein the pancreatic cancer is pancreatic ductal adenocarcinoma.

3. The pharmaceutical composition of claim 1 , wherein the dapanstril is administered by systemic administration.

4. The pharmaceutical composition of claim 3, wherein the dapanstril is administered orally.

5. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition reduces tumor volume or tumor weight.

6. The pharmaceutical composition of claim 1 , wherein the patient is not administered cytokine-induced killer cells.

7. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises a chemotherapeutic agent.

8. The pharmaceutical composition of claim 7, wherein the chemotherapeutic agent is gemcitabine.

9. 9. The pharmaceutical composition of claim 8, wherein the gemcitabine is administered intravenously.

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