Pharmaceutical rectal suppository compositions of 6-thioguanine, methods of treatment and / or methods of manufacture

Abstract

A rectal suppository is described. The rectal suppository contains 6-thioguanine, at least one stearin and at least one surfactant. Further or optionally, the rectal suppository comprises from about 1 mg to 9 mg of 6-thioguanine. Further or optionally, the present invention describes methods of preventing, treating and / or managing inflammatory bowel disease in a subject. The method comprises administering a rectal suppository to a subject in need thereof. Further or optionally, the present invention describes a method of making a rectal suppository. The present invention relates to a process for preparing a rectal suppository comprising the steps of (a) heating stearin to become liquid stearin, (b) adding at least one surfactant to the liquid stearin, (c) adding 6-thioguanine to the liquid stearin, (d) shaping stearin into a rectal suppository. The step (b) is optionally performed before or after the step (c).

Description

Background Technology

[0001] 6-Thioguanine (6-TG) is primarily used to treat acute non-lymphocytic leukemia. Oral immediate-release tablets marketed under the brand names Tabloid™ or Lanvis™ contain 40 mg of 6-thioguanine. Due to the high risk of hepatotoxicity, Tabloid™ and Lanvis™ are not recommended for maintenance therapy or similar long-term continuous treatment. Patients receiving 6-TG must be monitored for signs of hepatotoxicity. Furthermore, patients with genetic deficiencies in thiopurine methyltransferase (TPMT) or Nudix hydrolase 15 (NUDT15) face a higher risk of toxicity and typically require less than 10% of the standard 6-thioguanine dose.

[0002] Oral 6-thioguanine is marketed in the Netherlands under the brand name Thiosix™ for the treatment of inflammatory bowel disease (Crohn's disease or ulcerative colitis). The recommended daily dose is 0.3 mg / kg body weight, with a maximum dose of 25 mg / day. Each Thiosix™ tablet contains 10 mg or 20 mg and has a score to divide it into two doses. Tabloid™ and Lanvis™ are also used off-label for the treatment of inflammatory bowel disease in other countries, but are not registered for this purpose.

[0003] Crouwel F, Simsek M, van Doorn AS, Mulder CJJ, Buiter HJC, BarclayML, Florin TH, de Boer NK. “Rectally Administrated Thioguanine for Distal Ulcerative Colitis: A Multicenter Case Series”, Inflammatory Bowel Diseases, Vol. 29, No. 6, June 2023, pp. 1000-1004, describes the use of 20 mg thioguanine enemas and rectal suppositories for the treatment of proctitis and left-sided colitis. In Australia, these rectal suppositories are obtained on an individual prescription basis under a special access scheme applicable to situations where no licensed alternative pharmaceutical preparations are available. The 20 mg rectal suppositories are manufactured and dispensed by licensed dispensing pharmacies using thioguanine and excipients in accordance with Good Manufacturing Practices (GMP). The excipients used in the rectal suppositories are not disclosed in this paper. However, one of the authors of this paper, Timothy Florin (University of Queensland), has confirmed that the only ingredients in the rectal suppository are 6-thioguanine and cocoa butter. The 6-thioguanine is sieved through a 40# sieve before being mixed with the molten cocoa butter. A 40# sieve corresponds to 400 μm, therefore the particle size of the thioguanine is no larger than 400 μm.

[0004] References to any prior art in this specification do not imply an acknowledgment or implication that such prior art constitutes part of common general knowledge in any jurisdiction, nor do they imply that such prior art can be reasonably expected by a person skilled in the art to be understood, regarded as relevant, and / or combined with other prior art. Summary of the Invention

[0005] In a first aspect, a rectal suppository is provided, comprising: 6-Thioguanine, At least one stearin, and At least one surfactant.

[0006] In one aspect, a rectal suppository is provided that contains approximately 1 mg to 9 mg of 6-thioguanine.

[0007] In one aspect, a rectal suppository is provided, which contains about 1 mg to 9 mg of 6-thioguanine and at least one stearin.

[0008] In one aspect, a rectal suppository is provided, comprising about 1 mg to 9 mg of 6-thioguanine, at least one stearin, and at least one surfactant.

[0009] In at least one instance, the suppository contains about 1 mg to 30 mg of 6-thioguanine. In at least one instance, the suppository contains about 3 mg to 25 mg of 6-thioguanine. In at least one instance, the suppository contains about 5 mg to 20 mg of 6-thioguanine.

[0010] In at least one instance, the suppository contains about 8 mg to 30 mg of 6-thioguanine. In at least one instance, the suppository contains about 8 mg to 25 mg of 6-thioguanine. In at least one instance, the suppository contains about 10 mg to 20 mg of 6-thioguanine.

[0011] In at least one instance, the suppository contains about 15 mg to 30 mg of 6-thioguanine. In at least one instance, the suppository contains about 17 mg to 25 mg of 6-thioguanine. In at least one instance, the suppository contains about 20 mg of 6-thioguanine.

[0012] In at least one instance, the suppository contains about 6 mg to 15 mg of 6-thioguanine. In at least one instance, the suppository contains about 8 mg to 12 mg of 6-thioguanine. In at least one instance, the suppository contains about 10 mg of 6-thioguanine.

[0013] In at least one instance, the suppository contains about 1 mg to 9 mg of 6-thioguanine. In at least one instance, the suppository contains about 2 mg to 9 mg of 6-thioguanine. In at least one instance, the suppository contains about 3 mg to 9 mg of 6-thioguanine. In at least one instance, the suppository contains about 3 mg to 7 mg of 6-thioguanine. In at least one instance, the suppository contains about 4 mg to 6 mg of 6-thioguanine. In at least one instance, the suppository contains about 5 mg of 6-thioguanine.

[0014] In at least one instance, the suppository contains about 0.05% w / w to 5% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.05% w / w to 3% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.05% w / w to 2.5% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.05% w / w to 2% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.2% w / w to 1.5% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.3% w / w to 1.1% w / w 6-thioguanine.

[0015] In at least one instance, the suppository contains about 0.4% w / w to 2% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.4% w / w to 1.5% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.5% w / w to 1.1% w / w 6-thioguanine.

[0016] In at least one instance, the suppository contains about 0.6% w / w to 2% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.7% w / w to 1.5% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.8% w / w to 1.1% w / w 6-thioguanine. In at least one instance, the suppository contains about 1.1% w / w 6-thioguanine.

[0017] In at least one instance, the suppository contains about 0.4% w / w to 1.0% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.4% w / w to 0.9% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.4% w / w to 0.8% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.5% w / w 6-thioguanine.

[0018] In at least one instance, the suppository contains about 0.05% w / w to 0.4% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.1% w / w to 0.4% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.2% w / w to 0.4% w / w 6-thioguanine. In at least one instance, the suppository contains about 0.3% w / w 6-thioguanine.

[0019] In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 100 μm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 80 μm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 60 μm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 40 μm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 30 μm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of less than about 20 μm.

[0020] In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 100 µm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 80 µm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 60 µm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 40 µm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 30 µm. In at least one instance, the 6-thioguanine in the suppository has a particle size distribution with a D50 of about 1 μm to 20 µm.

[0021] In at least one instance, the suppository contains at least one type of stearin.

[0022] In at least one instance, the suppository contains approximately 80% w / w to 99.9% w / w stearin. In at least one instance, the suppository contains approximately 85% w / w to 99.5% w / w stearin. In at least one instance, the suppository contains approximately 90% w / w to 99.5% w / w stearin. In at least one instance, the suppository contains approximately 95% w / w to 99.5% w / w stearin.

[0023] In at least one instance, stearin comprises monoglycerides, diglycerides, and / or triglycerides of C10 to C18 fatty acids.

[0024] In at least one instance, stearin comprises monoglycerides, diglycerides, and triglycerides of C10 to C18 fatty acids.

[0025] In at least one instance, stearin comprises monoglycerides, diglycerides, and triglycerides of C10 to C18 fatty acids, with triglycerides being the predominant component.

[0026] In at least one instance, stearin contains less than about 10 mol% of one or more unsaturated fatty acids. In at least one instance, stearin contains less than about 5 mol% of one or more unsaturated fatty acids. In at least one instance, stearin contains less than about 2 mol% of one or more unsaturated fatty acids. In at least one instance, stearin contains less than about 1 mol% of one or more unsaturated fatty acids.

[0027] In at least one instance, the ester was prepared by transesterification of hydrogenated palm oil and hydrogenated palm kernel oil.

[0028] In at least one instance, the hydroxyl value of stearin is less than 40 mg KOH / g. In at least one instance, the hydroxyl value of stearin is from about 1 mg to 35 mg KOH / g. In at least one instance, the hydroxyl value of stearin is from about 1 mg to 30 mg KOH / g.

[0029] In at least one instance, the melting range of stearin is about 34°C to 38°C. In at least one instance, the melting range of stearin is about 34°C to 36°C.

[0030] In at least one instance, the suppository contains at least one surfactant.

[0031] In at least one instance, the surfactant has a hydrophilic-lipophilic balance (HLB) value equal to or greater than about 10.

[0032] In at least one instance, the surfactant is nonionic.

[0033] In at least one instance, the surfactant is selected from any one or more of the group consisting of polysorbate, polyoxyethylene derivatives of natural or hydrogenated vegetable oils, alkyl / dialkyl sulfates, sulfonates or sulfosuccinates, polyoxyethylene fatty acid esters, phospholipids, transesterification products of natural vegetable oil triglycerides and polyalkylene polyols, sorbitan fatty acid esters, pentaerythritol fatty acid esters, polyoxyethylene glycol alkyl ethers and / or polyoxyethylene glycol alkyl esters, sucrose esters, ethoxylated fatty alcohols, and fatty acid salts.

[0034] In at least one instance, the surfactant had a melting point below 35°C.

[0035] In at least one instance, the surfactant is polysorbate 80.

[0036] In at least one instance, the suppository contains about 0.1% w / w to 20% w / w of a surfactant. In at least one instance, the suppository contains about 0.1% w / w to 15% w / w of a surfactant. In at least one instance, the suppository contains about 0.1% w / w to 10% w / w of a surfactant. In at least one instance, the suppository contains about 0.1% w / w to 5% w / w of a surfactant. In at least one instance, the suppository contains about 0.2% w / w to 3% w / w of a surfactant. In at least one instance, the suppository contains about 0.4% w / w to 3% w / w of a surfactant. In at least one instance, the suppository contains about 0.5% w / w to 3% w / w of a surfactant. In at least one instance, the suppository contains about 0.5% w / w to 2% w / w of a surfactant.

[0037] In at least one instance, the suppository contains about 0.1% w / w to 5% w / w of polysorbate 80. In at least one instance, the suppository contains about 0.2% w / w to 3% w / w of polysorbate 80. In at least one instance, the suppository contains about 0.4% w / w to 3% w / w of polysorbate 80. In at least one instance, the suppository contains about 0.5% w / w to 3% w / w of polysorbate 80. In at least one instance, the suppository contains about 0.5% w / w to 2% w / w of polysorbate 80. In at least one instance, the suppository contains about 0.5% w / w to 1.5% w / w of polysorbate 80.

[0038] In at least one instance, the suppository contains a suspending agent.

[0039] In at least one instance, the suspending agent is present in an amount of about 0.05% w / w to 10% w / w, or about 0.1% w / w to 5% w / w, or about 0.1% w / w to 4% w / w, or about 0.1% w / w to 3% w / w, or about 0.1% w / w to 2% w / w, or about 0.1% w / w to 1% w / w.

[0040] In at least one instance, the suspending agent is selected from one or more of the following: silica, clay (including purified and / or refined clay), aluminum monostearate, and magnesium stearate.

[0041] In at least one instance, the clay is selected from one or more of kaolin, palygorskite, montmorillonite, sylvite, bentonite, halite, and magnesium aluminum silicate.

[0042] In at least one instance, the suspending agent includes silica.

[0043] In at least one instance, the silica is fumed silica and / or colloidal silica.

[0044] In at least one instance, the amount of silica present is about 0.05% w / w to 10% w / w, or about 0.1% w / w to 5% w / w, or about 0.1% w / w to 4% w / w, or about 0.1% w / w to 3% w / w, or about 0.1% w / w to 2% w / w, or about 0.1% w / w to 1% w / w.

[0045] In at least one instance, the suppository contains an antioxidant.

[0046] In at least one instance, the antioxidant is selected from butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, tocopherols (e.g., α-tocopherol (vitamin E)), propionic acid, sodium nitrate, sodium nitrite, anthocyanins, citric acid (e.g., citric acid monohydrate), and combinations thereof.

[0047] In at least one instance, the antioxidant includes butylated hydroxyanisole (BHA).

[0048] In at least one instance, antioxidants include vitamin E.

[0049] In at least one instance, the antioxidants included butylated hydroxyanisole (BHA) and vitamin E.

[0050] In at least one instance, the antioxidant is present in an amount of about 0.0003% w / w to 8% w / w, or about 0.003% w / w to 4% w / w, or about 0.003% w / w to 2.5% w / w.

[0051] In at least one instance, the suppository comprises: about 1 mg to 30 mg of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0052] In at least one instance, the suppository comprises: about 8 mg to 30 mg of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0053] In at least one instance, the suppository comprises: about 15 mg to 30 mg of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0054] In at least one instance, the suppository comprises: about 6 mg to 15 mg of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0055] In at least one instance, the suppository comprises: about 1 mg to 9 mg of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0056] In at least one instance, the suppository comprises: about 0.05% w / w to 5% w / w of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0057] In at least one instance, the suppository comprises: about 0.05% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0058] In at least one instance, the suppository comprises: about 0.4% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.5% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0059] In at least one instance, the suppository comprises: about 0.6% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.3% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0060] In at least one instance, the suppository comprises: about 0.4% w / w to 1% w / w of 6-thioguanine, about 80% w / w to 99.5% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0061] In at least one instance, the suppository comprises: about 0.05% w / w to 0.4% w / w of 6-thioguanine, about 80% w / w to 99.9% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0062] In at least one instance, the suppository comprises: about 0.05% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.8% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, about 0.05% w / w to 10% w / w of at least one suspending agent, and optionally at least one antioxidant.

[0063] In at least one instance, the suppository comprises: about 0.4% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.5% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, about 0.05% w / w to 10% w / w of at least one suspending agent, and optionally at least one antioxidant.

[0064] In at least one instance, the suppository comprises: about 0.6% w / w to 2% w / w of 6-thioguanine, about 80% w / w to 99.3% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, about 0.05% w / w to 10% w / w of at least one suspending agent, and optionally at least one antioxidant.

[0065] In at least one instance, the suppository comprises: about 0.4% w / w to 1% w / w of 6-thioguanine, about 80% w / w to 99.5% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, about 0.05% w / w to 10% w / w of at least one suspending agent, and optionally at least one antioxidant.

[0066] In at least one instance, the suppository comprises: about 0.05% w / w to 0.4% w / w of 6-thioguanine, about 80% w / w to 99.8% w / w of stearin, about 0.1% w / w to 20% w / w of at least one surfactant, about 0.05% w / w to 10% w / w of at least one suspending agent, and optionally at least one antioxidant.

[0067] In at least one instance, the total weight of the suppository was approximately 800 mg to 3000 mg. In at least one instance, the total weight of the suppository was approximately 1000 mg to 3000 mg. In at least one instance, the total weight of the suppository was approximately 1200 mg to 3000 mg. In at least one instance, the total weight of the suppository was approximately 1500 mg to 3000 mg. In at least one instance, the total weight of the suppository was approximately 1800 mg to 2200 mg. In at least one instance, the total weight of the suppository was approximately 1800 mg to 2000 mg. In at least one instance, the total weight of the suppository was approximately 1000 mg to 2500 mg. In at least one instance, the total weight of the suppository was approximately 1000 mg to 2000 mg.

[0068] In at least one instance, the suppository has a melting range of about 33°C to 38°C. In at least one instance, the suppository has a melting range of about 33°C to 37°C. In at least one instance, the suppository has a melting range of about 33°C to 36°C.

[0069] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which substantially corresponds to approximately 40% to 100%, or approximately 50% to 100%, or approximately 60% to 100%, or approximately 70% to 100% of 6-thioguanine when the suppository was placed in the dissolution medium for approximately 20 minutes.

[0070] In at least one instance, the suppository exhibits the following release of 6-thioguanine in an in vitro dissolution medium, which substantially corresponds to approximately 40% to 100%, or approximately 50% to 100%, or approximately 60% to 100%, or approximately 70% to 100%, or approximately 80% to 100% of 6-thioguanine when the suppository is placed in the dissolution medium for approximately 30 minutes.

[0071] In at least one instance, the suppository exhibited the following release of 6-thioguanine in the in vitro dissolution medium, which substantially corresponds to approximately 80% to 100%, or approximately 85% to 100%, or approximately 90% to 100% of the 6-thioguanine when the suppository was placed in the dissolution medium for approximately 60 minutes.

[0072] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which substantially corresponds to approximately 40%, or not less than approximately 60%, or not less than approximately 80% of the 6-thioguanine released approximately 60 minutes after the suppository was placed in the dissolution medium.

[0073] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which essentially corresponds to: (a) When the suppository is placed in the dissolution medium for about 20 minutes, about 20% to 100% of 6-thioguanine is released.

[0074] (b) When the suppository is placed in the dissolution medium for about 60 minutes, it shall contain not less than about 40% of 6-thioguanine.

[0075] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which essentially corresponds to: (a) When the suppository is placed in the dissolution medium for about 20 minutes, about 30% to 100% of 6-thioguanine is released.

[0076] (b) When the suppository is placed in the dissolution medium for about 60 minutes, it shall contain not less than about 60% of 6-thioguanine.

[0077] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which essentially corresponds to: (a) When the suppository is placed in the dissolution medium for about 20 minutes, about 40% to 100% of 6-thioguanine is released.

[0078] (b) When the suppository is placed in the dissolution medium for about 60 minutes, it shall contain no less than about 80% of 6-thioguanine.

[0079] In at least one instance, the suppository exhibited the following 6-thioguanine release in the in vitro dissolution medium, which essentially corresponds to: (a) When the suppository is placed in the dissolution medium for about 20 minutes, about 60% to 100% of 6-thioguanine is released.

[0080] (b) When the suppository is placed in the dissolution medium for about 60 minutes, it shall contain no less than about 80% of 6-thioguanine.

[0081] In at least one instance, the average 6-TGN provided by the suppository was less than approximately 150 pmol / 8 x 10⁻⁶ hours after suppository administration. 8 RBC.

[0082] In at least one instance, the average 6-TGN provided by the suppository was less than approximately 100 pmol / 8 x 10⁻⁶ hours after suppository administration. 8 RBC.

[0083] In at least one instance, 6 hours after suppository administration, the average 6-TGN provided by the suppository was less than approximately 75 pmol / 8 x 10⁻⁶. 8 RBC.

[0084] In at least one instance, the average 6-TGN provided by the suppository was less than approximately 60 pmol / 8 x 10⁻⁶ hours after suppository administration. 8 RBC.

[0085] In at least one instance, the average 6-TGN provided by the suppository was less than approximately 30 pmol / 8 x 10⁻⁶ hours after suppository administration. 8 RBC.

[0086] In at least one instance, 6 hours after suppository administration, the average 6-TGN provided by the suppository was less than approximately 25 pmol / 8 x 10⁻⁶. 8 RBC.

[0087] In at least one instance, the average 6-TGN provided by the suppository was less than approximately 20 pmol / 8 x 10⁻⁶ hours after suppository administration. 8 RBC.

[0088] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided lower levels of 6-TGN and / or 6-TG in the subject's breast milk.

[0089] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with 6-TGN and / or 6-TG levels of 15 ng / mL or less.

[0090] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with 6-TGN and / or 6-TG levels of 10 ng / mL or less.

[0091] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with 6-TGN and / or 6-TG levels of 5 ng / mL or less.

[0092] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with 6-TGN and / or 6-TG levels of 2.5 ng / mL or less.

[0093] In at least one instance, when the suppository was administered to the subject once daily, the levels of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository were undetectable.

[0094] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 15 ng / mL or less.

[0095] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 10 ng / mL or less.

[0096] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 5 ng / mL or less.

[0097] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 2.5 ng / mL or less.

[0098] In at least one instance, when suppositories were administered to subjects once daily for at least 28 days, the levels of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppositories were undetectable.

[0099] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided low levels of 6-TGN and / or 6-TG in the subject's breast milk, wherein the suppository contained approximately 1 mg to 9 mg of 6-thioguanine.

[0100] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with a level of 6-TGN and / or 6-TG of 15 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0101] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with a level of 6-TGN and / or 6-TG of 10 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0102] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with a level of 6-TGN and / or 6-TG of 5 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0103] In at least one instance, when the suppository was administered to the subject once daily, the suppository provided the subject's breast milk with a level of 6-TGN and / or 6-TG of 2.5 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0104] In at least one instance, when the suppository was administered to the subject once daily, the levels of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository were undetectable, wherein the suppository contained approximately 1 mg to 9 mg of 6-thioguanine.

[0105] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 15 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0106] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 10 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0107] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 5 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0108] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the level of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository was 2.5 ng / mL or less, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0109] In at least one instance, when the suppository was administered to the subject once daily for at least 28 days, the levels of 6-TGN and / or 6-TG in the subject's breast milk provided by the suppository were undetectable, wherein the suppository contained about 1 mg to 9 mg of 6-thioguanine.

[0110] To avoid doubt, where the context permits, the above implementation methods and / or embodiments may be applied individually or in any combination of two or more of them to any one or more of the following aspects.

[0111] In one aspect, a method is provided for preventing, treating, and / or managing inflammatory bowel disease in a subject, the method comprising administering the rectal suppository described in the first aspect to a subject in need.

[0112] In one aspect, a method for preventing, treating, and / or controlling inflammatory bowel disease in a subject is provided, the method comprising administering approximately 1 mg to 9 mg of 6-thioguanine in the form of a rectal suppository to the subject's rectum.

[0113] In one aspect, the use of the rectal suppository containing 6-thioguanine described in the first aspect in the manufacture of a medicament for the prevention, treatment and / or control of inflammatory bowel disease is provided.

[0114] In one aspect, the use of rectal suppositories containing 6-thioguanine in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease is provided, wherein the suppository contains about 1 mg to 9 mg of 6-thioguanine.

[0115] In one aspect, a rectal suppository containing 6-thioguanine as described in the first aspect for the prevention, treatment and / or control of inflammatory bowel disease is provided.

[0116] In one aspect, a rectal suppository containing 6-thioguanine is provided for the prevention, treatment, and / or control of inflammatory bowel disease, wherein the suppository contains about 1 mg to 9 mg of 6-thioguanine.

[0117] In one aspect, the rectal suppository containing 6-thioguanine described in the first aspect is provided for use in the prevention, treatment, and / or control of inflammatory bowel disease.

[0118] In one aspect, the use of rectal suppositories containing 6-thioguanine for the prevention, treatment, and / or control of inflammatory bowel disease is provided, wherein the suppository contains about 1 mg to 9 mg of 6-thioguanine.

[0119] In at least one instance, inflammatory bowel disease involved the rectum.

[0120] In at least one instance, inflammatory bowel disease is either ulcerative colitis or Crohn's disease that involves the rectum.

[0121] In at least one instance, ulcerative colitis is ulcerative proctitis, rectosigmoid colitis, and / or left colitis.

[0122] In at least one instance, the suppository was administered to the subject once or twice daily.

[0123] In at least one instance, the suppository was administered to the subject once daily.

[0124] In at least one instance, the suppository is formulated for administration to a patient once daily or twice daily.

[0125] In at least one instance, the suppository is formulated for administration to a patient once daily.

[0126] In at least one instance, the suppository provided a dose of 6-thioguanine of about 0.01 to 0.4 mg / kg body weight.

[0127] In at least one instance, the suppository provided a daily dose of 6-thioguanine of approximately 0.01 to 0.4 mg / kg body weight.

[0128] In at least one instance, the suppository provided a dose of 6-thioguanine of about 0.1 to 0.4 mg / kg body weight.

[0129] In at least one instance, the suppository provided a daily dose of 6-thioguanine of approximately 0.1 to 0.4 mg / kg body weight.

[0130] In at least one instance, the suppository provides a dose of 6-thioguanine of about 0.01 to 0.1 mg / kg body weight. In at least one instance, the suppository provides a dose of 6-thioguanine of 0.01 to 0.09 mg / kg body weight. In at least one instance, the suppository provides a dose of 6-thioguanine of 0.02 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a dose of 6-thioguanine of 0.03 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a dose of 6-thioguanine of 0.04 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a dose of 6-thioguanine of 0.05 to 0.08 mg / kg body weight.

[0131] In at least one instance, the suppository provides a daily dose of 6-thioguanine of about 0.01 to 0.1 mg / kg body weight. In at least one instance, the suppository provides a daily dose of 6-thioguanine of 0.01 to 0.09 mg / kg body weight. In at least one instance, the suppository provides a daily dose of 6-thioguanine of 0.02 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a daily dose of 6-thioguanine of 0.03 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a daily dose of 6-thioguanine of 0.04 to 0.08 mg / kg body weight. In at least one instance, the suppository provides a daily dose of 6-thioguanine of 0.05 to 0.08 mg / kg body weight.

[0132] In at least one instance, the suppository provided a clinical response in the subject within a period of less than 8 weeks after the subject began receiving the suppository once daily.

[0133] In at least one instance, the suppository provided a clinical response in the subject within a period of less than 6 weeks after the subject began receiving the suppository once daily.

[0134] In at least one instance, the suppository provided a clinical response in the subject within 30 days of the start of once-daily administration.

[0135] In at least one instance, the suppository provided a clinical response in the subject within a period of less than 22 days after the subject began receiving the suppository once daily.

[0136] In at least one instance, the suppository provided a clinical response in the subject within 8 days of the start of once-daily administration.

[0137] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0138] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0139] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0140] In at least one instance, within 6 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0141] In at least one instance, within 6 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0142] In at least one instance, within 6 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0143] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0144] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0145] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0146] In at least one instance, within 22 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0147] In at least one instance, within 22 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0148] In at least one instance, within 22 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0149] In at least one instance, within 15 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0150] In at least one instance, within 15 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0151] In at least one instance, within 15 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0152] In at least one instance, within 8 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2 points.

[0153] In at least one instance, within 8 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 3 points.

[0154] In at least one instance, within 8 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 4 points.

[0155] In at least one instance, the suppository provided relief in the subject for less than 8 weeks after the subject began receiving the suppository once daily.

[0156] In at least one instance, the suppository provided relief in the subject for less than 6 weeks after the subject began receiving the suppository once daily.

[0157] In at least one instance, the suppository provided relief in the subject within 30 days of the start of once-daily administration.

[0158] In at least one instance, the suppository provided relief in the subject within 22 days of the start of once-daily administration.

[0159] In at least one instance, the suppository provided relief in the subject within 15 days of the start of once-daily administration.

[0160] In at least one instance, the suppository provided relief in the subject within 8 days of the start of once-daily administration.

[0161] In at least one instance, the suppository provided relief in the subject for a period of less than 8 weeks after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0162] In at least one instance, the suppository provided relief in the subject for a period of less than 6 weeks after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0163] In at least one instance, the suppository provided relief in the subject for a period of less than 30 days after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0164] In at least one instance, the suppository provided relief in the subject for a period of less than 22 days after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0165] In at least one instance, the suppository provided relief in the subject for a period of less than 15 days after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0166] In at least one instance, the suppository provided relief in the subject for a period of less than 8 days after the subject began receiving the suppository once daily, wherein the suppository contained approximately 5 mg of 6-thioguanine.

[0167] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0168] In at least one instance, within 6 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0169] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0170] In at least one instance, within 22 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0171] In at least one instance, within 15 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0172] In at least one instance, within 8 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 to equal to or less than 2, with a reduction of greater than or equal to 3.

[0173] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Mayo Endoscopy Score (Mayo ES) by at least 1 point.

[0174] In at least one instance, within a period of less than 6 weeks after the subject began receiving suppositories once daily, the suppositories reduced the subject’s Mayo Endoscopy Score (Mayo ES) by at least 1 point.

[0175] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Mayo Endoscopy Score (Mayo ES) by at least 1 point.

[0176] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Mayo Endoscopy Score (Mayo ES) by at least 2 points.

[0177] In at least one instance, within a period of less than 6 weeks after the subject began receiving suppositories once daily, the suppositories reduced the subject’s Mayo Endoscopy Score (Mayo ES) by at least 2 points.

[0178] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Mayo Endoscopy Score (Mayo ES) by at least 2 points.

[0179] In at least one instance, within a period of less than 8 weeks after the subject began receiving suppositories once daily, the suppositories reduced the subject’s Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 1 point.

[0180] In at least one instance, within a period of less than 6 weeks after the subject began receiving suppositories once daily, the suppositories reduced the subject’s Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 1 point.

[0181] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 1 point.

[0182] In at least one instance, within 8 weeks of starting once-daily suppository administration to the subject, the suppository reduced the subject's Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 2 points.

[0183] In at least one instance, within a period of less than 6 weeks after the subject began receiving suppositories once daily, the suppositories reduced the subject’s Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 2 points.

[0184] In at least one instance, within 30 days of starting once-daily suppository administration to the subject, the suppository reduced the subject's Ulcerative Colitis Endoscopic Severity Index (UCEIS) by at least 2 points.

[0185] In at least one instance, additional inflammatory bowel disease treatment was administered concurrently with or at the same time as the administration of the rectal suppository.

[0186] In at least one instance, the additional inflammatory bowel disease treatment is selected from any one or more of 5-aminosalicylates (5-ASA, such as mesalazine, olsalazine, balsalazine, sulfasalazine), corticosteroids (such as prednisone), and biologics.

[0187] In at least one instance, other treatments for the symptoms of inflammatory bowel disease were administered concurrently with or during the same time period as the administration of rectal suppositories.

[0188] In at least one instance, other treatments for the symptoms of inflammatory bowel disease were selected from laxatives, antidiarrheals, and analgesics.

[0189] In one aspect, a method for manufacturing a rectal suppository is provided, the method comprising the following steps: (a) Heating stearin to turn it into liquid stearin. (b) Add at least one surfactant to the liquid stearin. (c) Add 6-thioguanine to the liquid stearin. (d) Form the stearin into a rectal suppository. Step (b) may be performed before or after step (c).

[0190] In at least one instance, step (b) is performed before step (c).

[0191] In at least one instance, stearin was heated to approximately 40°C to 90°C.

[0192] In at least one instance, stearin was heated to approximately 40°C to 80°C.

[0193] In at least one instance, stearin was heated to approximately 40°C to 70°C.

[0194] In at least one instance, stearin was heated to approximately 40°C to 65°C.

[0195] In at least one instance, stearin was heated to approximately 40°C to 60°C.

[0196] In at least one instance, stearin was stirred or agitated while being heated.

[0197] In at least one instance, a top-mounted mixer was used to stir the stearin.

[0198] In at least one instance, the method further includes the step of adding at least one antioxidant to the liquid stearin.

[0199] In at least one instance, at least one antioxidant was added to liquid stearin along with a surfactant.

[0200] In at least one instance, 6-thioguanine was added to the liquid stearate as a portion of the liquid stearate.

[0201] In at least one instance, after adding at least one surfactant, a portion of the liquid stearin was removed from the liquid stearin.

[0202] In at least one instance, a portion of the liquid stearin was removed from the liquid stearin after the addition of at least one surfactant and at least one antioxidant.

[0203] In at least one instance, the method further includes homogenizing liquid stearin, surfactant, and 6-thioguanine.

[0204] In at least one instance, the homogenization includes milling liquid stearin, surfactant, and 6-thioguanine.

[0205] In at least one instance, the method further includes reducing the particle size of 6-thioguanine.

[0206] In at least one instance, the particle size was reduced by grinding 6-thioguanine. In at least one instance, the particle size was reduced by wet grinding of 6-thioguanine.

[0207] In at least one instance, the grinding was performed after 6-thioguanine was added to liquid stearin.

[0208] In at least one instance, the grinding is ball milling.

[0209] In at least one instance, the ball mill uses beads with a diameter of about 0.1 mm to 2 mm. In at least one instance, the ball mill uses beads with a diameter of about 1 mm to 1.5 mm. In at least one instance, the ball mill uses beads with a diameter of about 1.2 mm to 1.4 mm.

[0210] In at least one instance, the method further includes the step of adding at least one suspending agent to the liquid stearate.

[0211] In at least one instance, the at least one suspending agent was added to the liquid stearin prior to 6-thioguanine.

[0212] In at least one instance, the at least one suspending agent is added to the liquid stearate after the surfactant.

[0213] In at least one instance, stearin is molded into suppositories by transferring liquid stearin into a mold.

[0214] In at least one instance, the stearin was cooled to about 31°C to 40°C before being transferred to the mold.

[0215] In at least one instance, the stearin was cooled to about 33°C to 35°C before being transferred to the mold.

[0216] In at least one instance, the stearin was cooled to about 37°C to 40°C before being transferred to the mold.

[0217] In at least one instance, the stearin was cooled to about 15°C to 25°C after being transferred to the mold. In at least one instance, the stearin was cooled to about 17°C to 22°C after being transferred to the mold.

[0218] To avoid doubt, where the context permits, implementation methods and / or embodiments may be applied individually or in any combination of two or more of them to any one or more of the above aspects.

[0219] Other aspects of the invention, as well as other embodiments and / or examples of the aspects described in the foregoing paragraphs, will become apparent from the following description, which is given by way of example and with reference to the accompanying drawings. Attached Figure Description

[0220] This invention will be described in conjunction with the following drawings, wherein the same reference numerals denote the same elements, and wherein: Figure 1 This is a schematic diagram of a method for manufacturing rectal suppositories.

[0221] Figure 2 This is a schematic diagram illustrating an example of manufacturing rectal suppositories.

[0222] Figure 3 The dissolution graphs were used to compare rectal suppository formulations with different stearates with a comparative example of suppositories made from cocoa butter.

[0223] Figure 4 This is a graph comparing the dissolution of rectal suppository formulations with different surfactants.

[0224] Figure 5 This is a graph comparing the uniformity of rectal suppository formulations with different silica contents.

[0225] Figure 6A , 6B6C is a histogram of the particle size distribution of thioguanine from different batches. Figure 6D This is a histogram of placebo rectal suppository formulations containing silica. Figure 6E This is a bar chart of ground rectal suppository formulations. Figure 6F1 This is a histogram of an unground rectal suppository formulation. Figure 6F2 This is a bar chart of the ground rectal suppository formulation. The histogram shows the y-axis as the bulk density (%), ranging from 0 to 8 (scaled at 0, 2, 4, 6, and 8), and the x-axis as the particle size distribution (µm), ranging from 0.01 to 10000.0 (scaled at 0.01, 0.1, 1.0, 10.0, 100.0, 1000.0, and 10000.0).

[0226] Figure 7 This is a graph comparing the dissolution of rectal suppository formulations with different contents of polysorbate 80.

[0227] Figure 8 This is the chromatogram of 6-TG.

[0228] Terminology Definition

[0229] As used herein, the terms “active pharmaceutical ingredient” (“API”), “pharmaceutical agent”, “active agent”, or “active ingredient” are drugs or pharmaceutical preparations that can be used as disclosed herein and are intended for use in humans or animals to cure, alleviate, prevent, or diagnose symptoms of disease, discomfort, physical injury, or pathology; allow a state, condition, or function of the body or mind to be identified; replace an active substance produced by the body or body fluids of the human or animal; defend against, eliminate, or render harmless to pathogens, parasites, or exogenous substances, or affect a state, condition, or function of the body or mind. 6-Thioguanine is an example of an API.

[0230] When the amount used herein provides an effect in subjects, that amount is “effective.” As used herein, the term “effective amount” or “therapeutic effective amount” means an amount of a compound or composition that is sufficient to significantly induce a positive benefit, including the individual benefits disclosed herein or combinations thereof, but low enough to avoid serious side effects, i.e., providing a reasonable benefit-risk ratio within the reasonable judgment of a person skilled in the art.

[0231] As used herein, in the context of administering a therapy to a subject, the term “prevention” means the prevention or suppression of a recurrence or flare-up of a disease or condition in a subject by administering one therapy (e.g., a preventative or therapeutic agent) or a combination of therapies (e.g., a combination of preventative or therapeutic agents), said disease or condition including inflammatory bowel disease, ulcerative colitis, Crohn's disease, ulcerative proctitis, rectosigmoiditis and / or left colitis or symptoms thereof.

[0232] As used herein, in the context of administering a therapy to a subject, the term “treatment” means reducing or inhibiting the progression and / or duration of a disease or condition (e.g., inflammatory bowel disease, ulcerative colitis, Crohn's disease, ulcerative proctitis, rectosigmoiditis, and / or left colitis), reducing or improving the severity of a disease or condition (e.g., inflammatory bowel disease, ulcerative colitis, Crohn's disease, ulcerative proctitis, rectosigmoiditis, and / or left colitis), and / or improving one or more of its symptoms caused by the administration of one or more therapies.

[0233] As used herein, in the context of administering a therapy to a subject, the term “manage” means preventing or inhibiting the progression of a disease or condition in a subject by administering one therapy (e.g., a preventative or therapeutic agent) or a combination of therapies (e.g., a combination of preventative or therapeutic agents), said disease or condition including inflammatory bowel disease, ulcerative colitis, Crohn's disease, ulcerative proctitis, rectosigmoiditis and / or left colitis or symptoms thereof.

[0234] As used herein, the terms “subject” and “patient” are used interchangeably. As used herein, the term “patient” refers to an animal, preferably a mammal such as a nonprimate (e.g., a cow, pig, horse, cat, dog, rat, etc.) and a primate (e.g., a monkey and a human), most preferably a human. In at least one instance, the subject is a non-human animal, such as a farm animal (e.g., a horse, pig, or cow) or a pet (e.g., a dog or cat). In at least one instance, the subject is an elderly person. In at least one instance, the subject is an adult. In at least one instance, the subject is a child. In at least one instance, the subject is an infant.

[0235] As used in this article, the phrase “pharmaceutical acceptable” means approved by a federal or state regulatory agency or listed in the United States Pharmacopeia, the European Pharmacopeia, or other recognized pharmacopoeias for use in animals, and more specifically, for use in humans.

[0236] As used herein, the term “about” when used in conjunction with the said value or range has the meaning reasonably assigned to it by those skilled in the art, meaning slightly higher or slightly lower than the said value or range, for example, it may vary by a maximum of 10% of the said value or range, or even by a maximum of 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0237] As used herein, unless the context otherwise requires, the term "comprise" and variations thereof, such as "comprising," "comprises," and "comprised," are not intended to exclude additional additives, components, integers, or steps.

[0238] As used herein, the term "stearin" refers to one or more glycerides of one or more saturated fatty acids, or one or more glycerides of one or more saturated fatty acids that are substantially solid at room temperature (e.g., about 20°C), or one or more monoglycerides, diglycerides, and / or triglycerides of one or more C10 to C18 saturated fatty acids that are substantially solid at room temperature (e.g., about 20°C), or one or more monoglycerides, diglycerides, and / or triglycerides of one or more C10 to C18 saturated fatty acids that are substantially solid at room temperature (e.g., about 20°C), containing less than about 10% of one or more unsaturated fatty acids, or less than about 5% of one or more unsaturated fatty acids, or less than about 2% of one or more unsaturated fatty acids, or less than about 1% of one or more unsaturated fatty acids. The percentage of unsaturation is based on molar percentage. One or more monoglycerides, diglycerides, and / or triglycerides of fatty acids (e.g., fatty acids of one or more chain lengths forming the ester, such as those selected from C10 to C18) may be present. The term "stearin" does not include cocoa butter.

[0239] As used herein, the in vitro dissolution or release assay of 6-thioguanine can be performed under the following conditions: Apparatus I (mesh basket, 40 mesh), speed 100 rpm ± 4 rpm, 900 mL of 0.05 M phosphate buffer solution in each dish, pH 6.8 ± 0.05, 37.0 °C ± 0.5 °C.

[0240] As used herein, the term "rectal suppository" refers to a dosage form formulated for rectal administration, meaning that the rectal suppository has a shape and / or size suitable for rectal administration and includes at least one suitable pharmaceutically acceptable excipient. The suitable pharmaceutically acceptable excipient melts at body temperature or is water-soluble and / or water-miscible, thereby enabling dispersion in rectal fluid and delivery of the API to the rectal region. The suitable pharmaceutically acceptable excipient is preferably substantially non-toxic and / or non-irritating to the mucosa. Detailed Implementation

[0241] This article describes a rectal suppository containing 6-thioguanine, at least one stearin and at least one surfactant.

[0242] Further or alternatively, this article describes a rectal suppository containing 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0243] Further or alternatively, this article describes a method for the prevention, treatment, and / or control of inflammatory bowel disease, the method comprising administering the rectal suppository described herein to a subject in need.

[0244] Further or alternatively, this article describes a method for preventing, treating, and / or controlling inflammatory bowel disease, the method comprising administering a rectal suppository containing 6-thioguanine, at least one stearin, and at least one surfactant.

[0245] Further or alternatively, this article describes a method for preventing, treating, and / or controlling inflammatory bowel disease, the method comprising administering a rectal suppository containing 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0246] Further or alternatively, this article describes the use of the rectal suppositories containing 6-thioguanine described herein in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease.

[0247] Further or alternatively, this article describes the use of rectal suppositories containing 6-thioguanine, at least one stearin and at least one surfactant in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease.

[0248] Further or alternatively, this article describes the use of rectal suppositories in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease, said rectal suppositories containing 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent and optionally at least one antioxidant.

[0249] Further or alternatively, this article describes the rectal suppositories containing 6-thioguanine described herein for the prevention, treatment and / or control of inflammatory bowel disease.

[0250] Further or alternatively, this article describes the rectal suppositories containing 6-thioguanine, at least one stearin and at least one surfactant, for the prevention, treatment and / or control of inflammatory bowel disease.

[0251] Further or alternatively, this article describes a rectal suppository containing 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant for the prevention, treatment, and / or control of inflammatory bowel disease.

[0252] Further or alternatively, this article describes the use of the rectal suppositories containing 6-thioguanine described herein for the prevention, treatment and / or control of inflammatory bowel disease.

[0253] Further or alternatively, this document describes the use of the rectal suppositories described herein, containing 6-thioguanine, at least one stearin, and at least one surfactant, for the prevention, treatment, and / or control of inflammatory bowel disease.

[0254] Further or alternatively, this article describes the use of a rectal suppository containing 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant for the prevention, treatment, and / or control of inflammatory bowel disease.

[0255] Further or alternatively, this article describes a rectal suppository containing approximately 1 mg to 9 mg of 6-thioguanine.

[0256] Further or alternatively, this article describes a rectal suppository containing about 1 mg to 9 mg of 6-thioguanine and at least one stearin.

[0257] Further or alternatively, this article describes a rectal suppository containing approximately 1 mg to 9 mg of 6-thioguanine and pharmaceutically acceptable excipients.

[0258] Further or alternatively, this article describes a rectal suppository containing about 1 mg to 9 mg of 6-thioguanine, at least one stearin and at least one surfactant.

[0259] Further or alternatively, this article describes a rectal suppository comprising about 1 mg to 9 mg of 6-thioguanine, at least one stearin, at least one surfactant, optionally at least one suspending agent, and optionally at least one antioxidant.

[0260] Further or alternatively, this article describes a method for the prevention, treatment, and / or control of inflammatory bowel disease in a subject, the method comprising administering approximately 1 mg to 9 mg of 6-thioguanine in the form of a rectal suppository to the subject's rectum.

[0261] Further or alternatively, this article describes the use of rectal suppositories containing 6-thioguanine in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease, wherein said suppositories contain about 1 mg to 9 mg of 6-thioguanine.

[0262] Further or alternatively, this article describes a rectal suppository containing 6-thioguanine for the prevention, treatment and / or control of inflammatory bowel disease, wherein the suppository contains about 1 mg to 9 mg of 6-thioguanine.

[0263] Further or alternatively, this article describes the use of rectal suppositories containing 6-thioguanine for the prevention, treatment and / or control of inflammatory bowel disease, wherein said suppositories contain about 1 mg to 9 mg of 6-thioguanine.

[0264] Further or alternatively, this document describes a method for manufacturing a rectal suppository, the method comprising the steps of: (a) heating stearin to become liquid stearin, (b) adding at least one surfactant to the liquid stearin, (c) adding 6-thioguanine to the liquid stearin, and (d) molding the stearin into a rectal suppository, wherein step (b) is optionally performed before or after step (c).

[0265] 6-Thioguanine

[0266] The structure of 6-thioguanine (thioguanine, 6-TG, 2-amino-1H-purine-6(7H)-thione) is shown in the following formula:

[0267] It will be apparent to those skilled in the art that 6-TG can be obtained and / or used as a hydrate (e.g., heptahydrate or hemihydrate) or a salt. When considering the amount, percentage, or proportion of 6-thioguanine in a pharmaceutical composition, the mass of water in the hydrate or salt should not be included.

[0268] 6-Thioguanine is a member of the thiopurine class of drugs. Thiopurine is a prodrug that is converted in vivo into its active metabolite, thioguanine nucleotide (6-TGN). The primary 6-TGN is 6-thioguanine triphosphate (6-TGTP), followed by 6-TGMP or 6-TGDP. Other members of the thiopurine family are 6-mercaptopurine (6-MP) and azathiopurine (AZA). However, the metabolic pathways of 6-MP and AZA are believed to differ from those of 6-TG. 6-MP and AZA are believed to be converted into 6-TGN in vivo by certain cells, including hepatocytes and leukocytes. This conversion pathway is rate-limited by inosine monophosphate dehydrogenase (IMPDH), therefore 6-TGN takes several days to appear, several weeks to reach a steady state, and several months to achieve clinical efficacy. Conversely, 6-thioguanine is believed to be metabolized into 6-TGTP via a more direct metabolic pathway by hypoxanthine-guanine phosphoribosyltransferase (HPRT). This may provide a faster clinical onset of action, but it can also cause serious hepatotoxicity, leading to hepatic vascular complications such as sinus obstruction syndrome (SOS), venous occlusive disease (VOD), or nodular regenerative proliferative disease (NRH). These side effects are not clearly associated with 6-MP or AZA.

[0269] WO 2017 / 054042 (also published as US 2016 / 15764247) describes the finding that induced colitis in mice lacking the key enzyme HPRT can be rapidly and significantly reduced upon treatment with 6-thioguanine, despite the absence of the usual metabolic pathway in the liver and leukocytes that converts 6-thioguanine to its active metabolite. It is believed that 6-thioguanine is converted to 6-TGTP via HPRT in the intestinal resident flora and / or diseased mucosa or epithelium and resident leukocytes at the site of inflammation. Therefore, it is believed that 6-thioguanine can act locally without producing significant systemic concentrations.

[0270] Rectal suppositories may contain a single thiopurine or purine analogue compound. It is believed that only thiopurine, 6-thioguanine, exerts a local effect at the site of application.

[0271] Rectal suppositories contain a therapeutically effective amount of 6-thioguanine.

[0272] Rectal suppositories may contain approximately 1 mg to 30 mg, or approximately 3 mg to 25 mg, or approximately 5 mg to 20 mg of 6-thioguanine in each suppository.

[0273] Alternatively, rectal suppositories may contain approximately 8 mg to 30 mg, or approximately 8 mg to 25 mg, or approximately 10 mg to 20 mg of 6-thioguanine in each suppository. Rectal suppositories may contain approximately 15 mg to 30 mg, or approximately 17 mg to 25 mg, or approximately 20 mg of 6-thioguanine in each suppository.

[0274] Alternatively, rectal suppositories may contain approximately 6 mg to 15 mg, or approximately 8 mg to 12 mg, or approximately 10 mg of 6-thioguanine in each suppository.

[0275] Alternatively, rectal suppositories may contain approximately 1 mg to 9 mg, or approximately 2 mg to 9 mg, or approximately 3 mg to 9 mg, or approximately 3 mg to 7 mg, or approximately 4 mg to 6 mg, or approximately 5 mg of 6-thioguanine in each suppository.

[0276] Because 6-thioguanine is known to be toxic at higher levels, low doses of 6-thioguanine in rectal suppositories, and / or low systemic levels of 6-thioguanine (and / or its metabolites), are beneficial while maintaining therapeutic efficacy.

[0277] Rectal suppository formulations may contain about 0.05% w / w to 5% w / w, or about 0.05% w / w to 3% w / w, or about 0.05% w / w to 2.5% w / w, or about 0.05% w / w to 2% w / w, or about 0.2% w / w to 1.5% w / w, or about 0.3% w / w to 1.1% w / w of 6-thioguanine.

[0278] Alternatively, rectal suppository formulations may contain about 0.4% w / w to 2% w / w, or about 0.4% w / w to 1.5% w / w, or about 0.5% w / w to 1.1% w / w of 6-thioguanine.

[0279] Alternatively, rectal suppository formulations may contain about 0.6% w / w to 2% w / w, or about 0.7% w / w to 1.5% w / w, or about 0.8% w / w to 1.1% w / w, or about 1.1% w / w of 6-thioguanine.

[0280] Alternatively, the rectal suppository formulation may contain about 0.4% w / w to 1.0% w / w, or about 0.4% w / w to 0.9% w / w, or about 0.4% w / w to 0.8% w / w, or about 0.5% w / w of 6-thioguanine.

[0281] Alternatively, the rectal suppository formulation may contain about 0.05% w / w to 0.4% w / w, or about 0.1% w / w to 0.4% w / w, or about 0.2% w / w to 0.4% w / w of 6-thioguanine, or about 0.3% w / w of 6-thioguanine.

[0282] 6-Thioguanine can be substantially suspended in stearin, while 6-thioguanine is essentially insoluble in stearin.

[0283] Various techniques can be used to characterize the particle size of materials. Those skilled in the art will also understand that almost all of these techniques do not directly measure the actual particle size as if measuring something with a ruler, but rather measure a physical phenomenon and interpret it as an indication of particle size.

[0284] For measurements performed using laser diffraction instruments or equivalent methods known in the art, the term "median particle size" is defined as the median particle size determined based on the equivalent spherical particle volume. When the term median is used, it should be understood to describe the particle size that divides a group of particles in half, i.e., 50% of the group is larger or smaller than that particle size. The median particle size is commonly denoted as D50, D(0.50), D[v,0.5], or D[0.5] or similar values. As used herein, D50, D[v,0.5], D(0.50), or D[0.5] or similar values ​​should be considered as referring to "median particle size".

[0285] The term "Dx of particle size distribution" (or similar) refers to the x-th percentile of the distribution; therefore, D90 refers to the 90th percentile, D95 to the 95th percentile, and so on. For example, D90 can be written as D[v,0.9], D(0.90), or D[0.9] or similar. Regarding median particle size and Dx, the uppercase letter D and the lowercase letter d are used interchangeably and have the same meaning.

[0286] Particle size distribution (e.g., the particle size distribution of APIs and / or other components of the formulation) can be described by volume distribution at different volume percentages, for example, at 10%, 50%, and 90% of the volume.

[0287] A common way to describe particle size distribution measured by laser diffraction or equivalent methods known in the art is to describe the percentage of particles smaller than or larger than a specified particle size. The term "percentage less than" is also written as "%<", and it is defined as the percentage of particles in the particle size distribution, by volume, that are smaller than a specified particle size—for example, %<1000 nm. The term "percentage greater than" is also written as "%>", and it is defined as the percentage of particles in the particle size distribution, by volume, that are larger than a specified particle size—for example, %>1000 nm.

[0288] The D50 particle size distribution of 6-thioguanine in rectal suppositories can be less than about 100 μm, or less than about 80 μm, or less than about 60 μm, or less than about 40 μm, or less than about 30 μm, or less than about 20 μm.

[0289] The D50 particle size distribution of 6-thioguanine in rectal suppositories can be approximately 1 μm to 100 μm, or approximately 1 μm to 80 μm, or approximately 1 μm to 60 μm, or approximately 1 μm to 40 μm, or approximately 1 μm to 30 μm, or approximately 1 μm to 20 μm.

[0290] In particular, the suppository formulation can be milled during the manufacturing process. In this case, the D50 particle size distribution of 6-thioguanine in the rectal suppository can be less than about 50 μm, or 40 μm, or less than about 30 μm, or less than about 20 μm, or the D50 particle size distribution of 6-thioguanine in the rectal suppository can be from about 1 μm to 50 μm, or from about 1 μm to 40 μm, or from about 1 μm to 30 μm, or from about 1 μm to 20 μm.

[0291] The D50 particle size distribution (the D50 particle size distribution of all particles in the suppository) of a rectal suppository can be less than about 50 μm, or less than about 40 μm, or less than about 30 μm, or less than about 25 μm, or the D50 particle size distribution of a rectal suppository can be from about 1 μm to 50 μm, or about 1 μm to 40 μm, or about 1 μm to 30 μm, or about 10 μm to 30 μm. The D50 particle size distribution of a rectal suppository can be from about 1 μm to 40 μm.

[0292] stearin

[0293] Pharmaceutically acceptable stearates are commercially available. The term "stearate" in the name refers to its physical state at room temperature (e.g., 20°C). However, stearates are designed to melt between 33°C and 39°C. The melting range varies among different stearates. Examples are shown in Table 1.

[0294] Stearins are typically produced synthetically. For example, in the Suppocire series of stearins available from Gattefossé, products without the prefix "N" are prepared through transesterification of hydrogenated palm oil and hydrogenated palm kernel oil. The prefix "N" indicates stearins obtained through direct esterification of fatty acids with glycerol. "M" indicates a low hydroxyl value (<15). The Suppocire series offers fats with different melting points / melting ranges, from as low as 32.5°C to as high as >40°C. The letters in this series indicate the melting point / melting range, for example, AI < A < B < C < D.

[0295] The Witopsol series of stearins from IOI Oleochemical comprises glycerides of saturated vegetable fatty acids (primarily lauric acid). The raw material is purified coconut and palm kernel oil. After initial purification, the oil is cracked with water under high pressure and an acidic or alkaline catalyst to obtain its fatty acids and glycerol. The fatty acid mixture is catalytically hydrogenated, followed by fractional vacuum distillation to remove low molecular weight hexanoic acid, octanoic acid, and capric acid (C6-C10). The C12-C18 fatty acids are adjusted to the correct mixture for this stearin grade and esterified with glycerol. The fatty acid profile, the stoichiometry of the reaction mixture, and the reaction time and temperature determine the properties of the product, such as melting range, solid fat index, hardness, monoglyceride, diglyceride, triglyceride content (emulsifying / dispersing properties), and viscosity.

[0296] Some commercially available stearins also include additives (such as beeswax) to alter viscosity or consistency.

[0297] For comparison, cocoa butter (cocoa oil) is also considered an oily matrix used in rectal suppositories. However, cocoa butter is composed of a mixture of triglycerides, primarily consisting of the fatty acids oleic acid (C18:1), palmitic acid (C16:0), and stearic acid (C18:0). It is a natural product, therefore its composition varies, but the saturated fatty acid content is typically about 57% to 64%, and the unsaturated fatty acid content is about 36% to 43%. Cocoa butter has a melting range of about 30°C to 35°C, but as a polycrystalline compound, its melting point can change to about 25°C to 30°C if heated above 35°C.

[0298] Table 1: Examples of commercially available stearins

[0299] Rectal suppositories may contain approximately 80% w / w to 99.9% w / w stearin, or approximately 85% w / w to 99.5% w / w stearin, or approximately 90% w / w to 99.5% w / w stearin, or approximately 95% w / w to 99.5% w / w stearin, or approximately 80% w / w to 99% w / w stearin, or approximately 85% w / w to 99% w / w stearin, or approximately 90% w / w to 99% w / w stearin, or approximately 95% w / w to 99% w / w stearin.

[0300] Rectal suppository formulations may contain stearin, said stearin comprising monoglycerides, diglycerides, and / or triglycerides of C10 to C18 fatty acids. Stearin may comprise monoglycerides, diglycerides, and / or triglycerides of C10 to C18 fatty acids. Triglycerides may be the predominant component.

[0301] Rectal suppositories may contain triglycerides prepared by transesterification of hydrogenated palm oil and hydrogenated palm kernel oil.

[0302] Rectal suppositories may contain stearin with a hydroxyl value of less than 40 mg KOH / g, or a hydroxyl value of about 1 to 35 mg KOH / g, or a hydroxyl value of about 1 to 30 mg KOH / g. Stearin with a higher hydroxyl value is believed to have poorer chemical stability, such as being more prone to oxidation. The melting range of rectal suppositories may be about 34°C to 38°C, or about 34°C to 36°C (e.g., at 1 atmosphere).

[0303] Alternative suppository base

[0304] The rectal suppository contains at least one suitable pharmaceutically acceptable excipient that melts at body temperature (e.g., an oily matrix, such as stearin or cocoa butter / cocoa soybean oil, emulsified cocoa soybean oil); or is water-soluble and / or water-miscible, thereby enabling dispersion in rectal fluid (e.g., a hydrophilic matrix, such as polyethylene glycol (PEG) of various molecular weights and / or one or a mixture of fatty acid esters of polyethylene glycol), which enables the delivery of the API to the rectal region.

[0305] Rectal suppositories may contain approximately 1 mh to 9 mg of 6-thioguanine and pharmaceutically acceptable excipients selected from oily and hydrophilic matrices.

[0306] surfactants

[0307] Examples of commercially available surfactants and their hydrophilic-lipophilic balance (HLB) values ​​are shown in Table 2. The hydrophilic-lipophilic balance (HLB) is the weight percentage of hydrophilic groups to hydrophobic groups in a molecule. The most commonly used HLB scale is 1 to 20; however, in some cases, to accommodate outliers, such as sodium dodecyl sulfate (SLS) with an HLB value of approximately 40, the scale can be higher (e.g., 1 to 40).

[0308] Table 2: Examples of surfactants

[0309] As shown in Examples 2 and / or 5, in in vitro dissolution tests, surfactants increase the dissolution rate and / or the amount of dissolution. In vitro dissolution tests can be used to reflect the in vivo performance of 6-TG diffusion and dissolution from rectal suppositories.

[0310] Rectal suppositories may contain nonionic surfactants.

[0311] Rectal suppositories may contain surfactants with a hydrophilic-lipophilic balance (HLB) value equal to or greater than about 10.

[0312] Rectal suppositories may contain suitable surfactants, such as, but not limited to, polysorbate esters, polyoxyethylene derivatives of natural or hydrogenated vegetable oils (e.g., castor oil), alkyl / dialkyl sulfates, sulfonates or sulfosuccinates, polyoxyethylene fatty acid esters, phospholipids, transesterification products of natural vegetable oil triglycerides and polyalkylene polyols, sorbitan fatty acid esters, pentaerythritol fatty acid esters, polyoxyethylene glycol alkyl ethers and / or esters; sucrose esters, ethoxylated fatty alcohols, fatty acid salts, and mixtures thereof.

[0313] Examples of polysorbates (e.g., polyoxyethylene-sorbitan fatty acid esters, such as mono-, di- and trilaurate, palmitate, stearate and oleate) include polysorbate 80, polysorbate 65, polysorbate 60, polysorbate 40 and polysorbate 20.

[0314] An example of a polyoxyethylene derivative of a natural or hydrogenated vegetable oil (such as castor oil) is polyoxyethylene 35 castor oil.

[0315] Examples of alkyl / dialkyl sulfates, sulfonates, or sulfosuccinates include sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and disodium lauryl polyoxyethylene ether sulfosuccinate.

[0316] An example of a polyoxyethylene fatty acid ester is PEG 100 stearate.

[0317] An example of phospholipids is lecithin.

[0318] Examples of sorbitan fatty acid esters include sorbitan isostearate, sorbitan sesquioleate, sorbitan monooleate, sorbitan monostearate, sorbitan tripearate, sorbitan monopalmitate, and sorbitan monolaurate.

[0319] Examples of polyoxyethylene glycol alkyl ethers and / or esters include polyethylene glycol octadecyl ether and polyethylene glycol 12 cetearyl alcohol ether.

[0320] Examples of sucrose esters (such as sucrose fatty acid esters) include sucrose palmitate and sucrose stearate.

[0321] Examples of salts of fatty acids (such as oleic acid) include potassium oleate and sodium oleate.

[0322] Examples of specific surfactants that may be used include, but are not limited to, polyoxyethylene castor oil derivatives, such as polyoxyethylene glycerol triricinoleate polyoxyethylene 35 castor oil (e.g., CREMOPHOR® EL, available from BASF), and polyoxyethylene 40 hydrogenated castor oil (e.g., CREMOPHOR® RH40, available from BASF); monofatty acid esters of polyoxyethylene (20) sorbitan (e.g., polyoxyethylene (20) sorbitan monooleate (TWEEN® 80), polyoxyethylene (20) sorbitan monostearate (TWEEN® 60), polyoxyethylene (20) sorbitan monopalmitate (TWEEN® 40), and polyoxyethylene (20) sorbitan monolaurate (TWEEN® 20); polyoxyethylene glycol alkyl ethers and polyoxyethylene glycol alkyl esters (e.g., polyethylene glycol octadecyl ether (Brij S20, polyoxyethylene (20) stearyl ether)) and mixtures thereof.

[0323] Rectal suppositories may contain polysorbate 80.

[0324] Surfactants may be liquid at room temperature (e.g., about 20°C and 1 atmosphere) and / or have a melting point below 35°C (e.g., at 1 atmosphere).

[0325] Rectal suppositories may contain a surfactant in the form of approximately 0.1% w / w to 20% w / w, or approximately 0.1% w / w to 15% w / w, or approximately 0.1% w / w to 10% w / w, or approximately 0.1% w / w to 5% w / w, or approximately 0.1% w / w to 5% w / w, or approximately 0.2% w / w to 3% w / w, or approximately 0.4% w / w to 3% w / w, or approximately 0.5% w / w to 3% w / w, or approximately 0.5% w / w to 2% w / w.

[0326] Rectal suppositories may contain about 0.1% w / w to 5% w / w, or about 0.2% w / w to 3% w / w, or about 0.4% w / w to 3%, or about 0.5% w / w to 3% w / w, or about 0.5% w / w to 2% w / w, or about 0.5% w / w to 1.5% w / w polysorbate 80.

[0327] Suspending agent

[0328] Rectal suppositories may contain suspending agents.

[0329] Suspension agents can improve the homogeneity of a batch of rectal suppositories, for example, by reducing the variation in the amount of 6-thioguanine between individual suppositories within a batch and / or reducing the standard deviation of the amount of 6-thioguanine in individual suppositories within a batch. Suspension agents can also alter the viscosity of a rectal suppository formulation to provide suspension. In this context, suspending agents may also be referred to as viscosity modifiers.

[0330] As shown in Example 3, in a batch of units / suppositories prepared from a rectal suppository formulation, the addition of a suspending agent improved the uniformity of the amount of 6-thioguanine in each unit / suppository.

[0331] Rectal suppositories may contain a suspending agent of about 0.05% w / w to 10% w / w, or about 0.1% w / w to 5% w / w, or about 0.1% w / w to 4% w / w, or about 0.1% w / w to 3% w / w, or about 0.1% w / w to 2% w / w, or about 0.1% w / w to 1% w / w.

[0332] Rectal suppositories may contain a suspending agent selected from one or more of silica, clay (including purified and / or refined clay), aluminum monostearate, and magnesium stearate. The clay may be selected from one or more of kaolin, palygorskite, montmorillonite, sylvite, bentonite, rock salt, and magnesium aluminum silicate.

[0333] Suspension agents may include silica. Silica may be fumed silica and / or colloidal silica. Examples of commercially available silica-based suspending agents include the Aerosil™ product line from Evonik.

[0334] Rectal suppositories may contain approximately 0.05% w / w to 10% w / w, or approximately 0.1% w / w to 5% w / w, or approximately 0.1% w / w to 4% w / w, or approximately 0.1% w / w to 3% w / w, or approximately 0.1% w / w to 2% w / w, or approximately 0.1% w / w to 1% w / w of silica.

[0335] Alternative or additional methods for improving and / or controlling the uniformity of rectal suppositories in a batch may include controlling the temperature of the liquid stearin (e.g., controlling the viscosity of the stearin) and / or controlling the stirring of the liquid stearin and / or other methods.

[0336] Other excipients

[0337] Over time, stearin may oxidize. Rectal suppositories may contain antioxidants to mitigate the oxidation of stearin and / or other components. Examples of suitable antioxidants may include one or more of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, tocopherols (e.g., α-tocopherol (vitamin E)), propionic acid, sodium nitrate, sodium nitrite, anthocyanins, and citric acid (e.g., citrate monohydrate). Rectal suppositories may contain butylated hydroxyanisole (BHA) and / or vitamin E.

[0338] The antioxidant present in rectal suppositories may be from about 0.0003% w / w to 8% w / w, or from about 0.003% w / w to 4% w / w, or from about 0.003% w / w to 2.5% w / w.

[0339] treat

[0340] Rectal suppositories may be used for the prevention, treatment, and / or control of inflammatory bowel disease in subjects who require them. Inflammatory bowel disease preferably involves the rectum.

[0341] Inflammatory bowel disease can be ulcerative colitis or Crohn's disease that involves the rectum. For example, ulcerative colitis can be ulcerative proctitis, rectosigmoid colitis, and / or left colitis.

[0342] Rectal suppositories can be administered once or twice daily, or formulated for once or twice daily use.

[0343] Rectal suppositories are well-suited for rectal delivery, for example, their size and / or shape are conducive to administration and / or retention. Compared to enemas, rectal suppositories are generally easier for patients to administer and retain, thus improving treatment adherence and gaining greater patient preference. Compared to enemas, rectal suppositories have a smaller contact area with the digestive tract and / or are located lower, which makes them particularly suitable for the treatment of ulcerative proctitis. Suppositories are generally more concentrated than enemas, for example, in volume, thus allowing for the delivery of higher concentrations of active substances (e.g., 6-thioguanine) to local areas (e.g., the rectum). Compared to the same dose of an enema, rectal suppositories can result in lower levels of 6-TGN in RBCs, which can provide lower toxicity.

[0344] The total weight of the rectal suppository may be approximately 800 mg to 3000 mg, or approximately 1000 mg to 3000 mg, or approximately 1200 mg to 3000 mg, or approximately 1500 mg to 3000 mg, or approximately 1800 mg to 2200 mg, or approximately 1800 mg to 2000 mg, or approximately 1000 mg to 2500 mg, or approximately 1000 mg to 2000 mg.

[0345] Rectal suppositories can be substantially cylindrical, preferably having at least one rounded end. Alternatively, rectal suppositories can be torpedo-shaped, oval (including elongated oval), or conical (including truncated conical).

[0346] Rectal suppositories provide a single dose. This dose cannot be split, nor can multiple doses be administered at once. Therefore, the dose delivered to the subject is based on the amount of 6-thioguanine in the rectal suppository. The dose of the rectal suppository used can be selected based on the subject's weight, or, for a group, based on a weight range of the group. For example, the dose can be selected based on a weight range of 60 kg to 80 g; however, the weight range used can vary by region.

[0347] Rectal suppositories are available in doses (e.g., daily dose) of about 0.01 to 0.4 mg / kg body weight of 6-thioguanine or in doses (e.g., daily dose) of about 0.1 to 0.4 mg / kg body weight of 6-thioguanine.

[0348] Methods for the prevention, treatment, and / or control of inflammatory bowel disease in subjects may include the administration of a suppository containing as little as about 1 mg to 9 mg of 6-thioguanine to the rectum of the subject. Previously, the oral dose of 6-thioguanine was 40 mg tablets (about 2 mg / kg body weight / day) for the treatment of acute non-lymphocytic leukemia, or 10 mg or 20 mg tablets (0.3 mg / kg body weight / day) for the treatment of Crohn's disease or ulcerative colitis. Currently, there are no registered rectal products for 6-thioguanine. However, previous trials have used a single rectal dose of 20 mg (enema or suppository) (approximately 0.25 mg / kg to 0.33 mg / kg body weight) – see, for example, Crouwel F, Simsek M, van Doorn AS, Mulder CJJ, Buiter HJC, Barclay ML, Florin TH, de Boer NK. “Rectally Administrated Thioguanine for Distal Ulcerative Colitis: A Multicenter Case Series”, Inflammatory Bowel Diseases, Vol. 29, No. 6, June 2023, pp. 1000-1004. Therefore, it is surprising that low-dose rectal suppositories of approximately 1 mg to 9 mg of 6-thioguanine have proven effective.

[0349] Rectal suppositories can provide daily doses of 6-thioguanine as low as approximately 0.01 to 0.1 mg / kg body weight, or approximately 0.01 to 0.09 mg / kg body weight, or approximately 0.02 to 0.08 mg / kg body weight, or approximately 0.03 to 0.08 mg / kg body weight, or approximately 0.04 to 0.08 mg / kg body weight, or approximately 0.05 to 0.08 mg / kg body weight.

[0350] Although the terms "daily," "twice daily," "once a day," or "twice a day" are mentioned, this refers to the number of times the rectal suppository is administered within a day. Those skilled in the art will understand that rectal suppositories do not necessarily need to be administered daily; for example, they can be administered every 2, 3, 4, 5, 6, or 7 days (or other). For instance, rectal suppositories may not need to be administered daily, especially when used as a maintenance dose.

[0351] Other treatments for inflammatory bowel disease (IBD) or IBD symptom management can be administered at the same time or within the same timeframe as the rectal suppository. An example of simultaneous administration could be within one hour of suppository administration. An example of simultaneous timeframe could be if the suppository is administered once daily, twice daily, or every two days for one week, in which case other treatments are used within the same week. Examples of other IBD treatments include 5-aminosalicylic acid (5-ASA) (e.g., mesalazine, olsalazine, balsalazine, sulfasalazine), corticosteroids (e.g., prednisone), and / or biologics. Examples of IBD symptom management include laxatives, antidiarrheals, and analgesics. IBD is commonly associated with diarrhea; however, constipation can become a problem, particularly during treatment.

[0352] Six hours after administration of the rectal suppository, the suppository reduced the average level of 6-TGN by approximately 150 pmol / 8 x 10⁻⁶. 8 RBC, or below approximately 100 pmol / 8 x 10⁻⁶ 8 RBC, or below approximately 75 pmol / 8 x 10⁻⁶ 8 RBC, or below approximately 60 pmol / 8 x 10 8 RBC, or below approximately 30 pmol / 8 x 10 8 RBC, or below approximately 25 pmol / 8 x 10⁻⁶ 8 RBC, or below approximately 20 pmol / 8 x 10⁻⁶ 8 RBCs. However, lower 6-TGN levels in RBCs are preferred while still maintaining therapeutic efficacy. In particular, for rectal suppositories containing approximately 1 mg to 9 mg of 6-thioguanine, rectal suppositories can reduce the average 6-TGN level to below approximately 30 pmol / 8 x 10⁻⁶ hours after suppository administration. 8RBC, or below approximately 25 pmol / 8x10 8 RBC, or below approximately 20 pmol / 8 x 10⁻⁶ 8 RBC. 6-TGN levels above 450 pmol / 8 x 10⁻⁶ 8 RBC is associated with a high risk of leukopenia and myelotoxicity (Afrasyab Khan, Arvenia B Berahmana, Andrew S Day, Murray L Barclay, Michael Schultz, New Zealand Medical Association Journal (NZMJ), March 8, 2019, Vol. 132, No. 1491, ISSN 1175-8716).

[0353] Once the subject is in a steady state, 6-TGN or 6-TG levels can be measured, for example, if the subject has been taking suppositories at a once-daily dose for at least 2 weeks, or at least 4 weeks. "Steady state" refers to a situation where the drug concentration remains consistently within the therapeutic limit over a long period. The concentration at which the drug concentration remains consistently near its therapeutic limit is called the "steady-state concentration."

[0354] Rectal suppositories are considered safe for use during lactation. When administered once daily to a subject, the suppository provides low levels of 6-TGN and / or 6-TG in the subject's breast milk, for example, at or below 15 ng / mL, at or below 10 ng / mL, at or below 5 ng / mL, and at or below 2.5 ng / mL. Breast milk 6-TGN and / or 6-TG levels can be measured after administering the suppository once daily to the subject for at least 28 days. Breast milk 6-TGN levels can be measured at 2, 6, or 9 hours following administration of the rectal suppository. Specifically, the rectal suppository may contain approximately 1 mg to 9 mg of 6-thioguanine, or approximately 5 mg.

[0355] Once the rectal suppository is administered to the subject once daily, the suppository can provide a clinical response in the subject within a timeframe of less than 8 weeks, less than 6 weeks, less than 30 days, less than 22 days, or less than 8 days.

[0356] When the rectal suppository was administered to the subject once daily, the suppository reduced the subject's Simple Clinical Colitis Activity Index (SCCAI) score by more than 2, 3, or 4 points within 8 weeks, 6 weeks, 30 days, 22 days, 15 days, or 8 days, respectively.

[0357] Once once-daily administration of rectal suppositories to subjects is initiated, the suppositories can provide disease remission in subjects within a timeframe of less than 8 weeks, less than 6 weeks, less than 30 days, less than 15 days, or less than 8 days. Disease remission after the start of treatment can be defined as a decrease in the subject's Simple Clinical Colitis Activity Index (SCCAI) score from greater than 2 points to equal to or less than 2 points, or a decrease of greater than or equal to 3 points.

[0358] After initiating once-daily administration of rectal suppositories to subjects, the suppositories can reduce a subject's Mayo Endoscopy Score (Mayo ES) by at least 1 point or improve it by at least 2 points within a timeframe of 8 weeks, 6 weeks, or 30 days. For example, if the Mayo ES is measured before initiating once-daily treatment, and a reduction of 1 or 2 points or more is observed within a timeframe of 8 weeks, 6 weeks, or 30 days of treatment.

[0359] After initiating once-daily administration of rectal suppositories to subjects, the suppositories can reduce the subject's Ulcerative Colitis Endoscopic Severity Index (UCEIS) score by at least 1 point or at least 2 points within a timeframe of 8 weeks, 6 weeks, or 30 days. For example, if the UCEIS score is measured before initiating once-daily treatment, and a reduction of 1 or more points is observed within a timeframe of 8 weeks, 6 weeks, or 30 days of treatment.

[0360] Methods of manufacturing suppositories

[0361] This article describes a method for manufacturing a rectal suppository, the method comprising the steps of: (a) heating stearin to become liquid stearin, (b) adding at least one surfactant to the liquid stearin, (c) adding 6-thioguanine to the liquid stearin, and (d) molding the stearin into a rectal suppository, wherein step (b) is optionally performed before or after step (c).

[0362] Examples of stearin, surfactants, and 6-thioguanine are as described above.

[0363] Reference Figure 1 In step (i), the stearin is melted to obtain a liquid. The stearin may be heated to about 40°C to 90°C, or about 40°C to 80°C, or about 40°C to 70°C, about 40°C to 65°C, or about 40°C to 60°C, or about 20°C above the melting range of the stearin, and optionally stirred or agitated while heating, for example using a top-mounted stirrer.

[0364] Reference Figure 1The steps (ii) of adding at least one surfactant to the liquid stearin and (iii) of adding 6-thioguanine to the liquid stearin can be performed in any order (e.g., (ii) before (iii) or (iii) before (ii), or substantially simultaneously. During steps (ii) and (iii), the heating and / or stirring / agitation of step (i) may continue.

[0365] Step (ii), adding at least one surfactant to the liquid stearin, may be carried out prior to (iii) adding 6-thioguanine to the liquid stearin.

[0366] The method may also include the step of adding at least one antioxidant to the liquid stearin. The at least one antioxidant may be added to the liquid stearin together with a surfactant, for example, simultaneously or sequentially.

[0367] 6-Thioguanine can be added to liquid stearin in the form of a portion of the liquid stearin, for example, a portion of the liquid stearin can be removed from the liquid stearin after the addition of at least one surfactant and / or at least one antioxidant.

[0368] The method may further include the step of adding at least one suspending agent to the liquid stearin. The at least one suspending agent may optionally be added to the liquid stearin before 6-thioguanine. The at least one suspending agent may optionally be added to the liquid stearin before or after a surfactant, and optionally before or after an antioxidant.

[0369] The method may also include homogenizing the liquid stearin after adding 6-thioguanine. Homogenization may include milling and / or high-pressure homogenization. For example, wet milling may include wet milling of a stearin mixture containing 6-thioguanine and a surfactant, and optionally an antioxidant and / or a suspending agent. Examples of milling include ball milling. When using a ball mill, the beads may be about 0.1 mm to 2 mm, or about 1 mm to 1.5 mm, or about 1.2 mm to 1.4 mm.

[0370] The particle size of 6-thioguanine can vary. In particular, when 6-thioguanine has a large particle size (e.g., D50 of about 20 μm to 100 μm) or a large particle size distribution, the particle size can be reduced before incorporation into rectal suppositories (e.g., by dry milling, such as jet milling) or after mixing with liquid stearin (e.g., by wet milling).

[0371] Grinding can help homogenize stearin and 6-thioguanine (e.g., make the distribution of 6-thioguanine more consistent throughout the stearin) and / or reduce the particle size of 6-thioguanine particles (and / or particles of other components in the suppository), and / or reduce the particle size distribution of 6-thioguanine particles (and / or particles of other components in the suppository), for example, by reducing the number of larger 6-thioguanine particles.

[0372] Liquid stearin can be molded into rectal suppositories by transferring it into a mold. The stearin can be cooled to approximately 31°C to 40°C, for example, approximately 33°C to 35°C, before being transferred to the mold. Alternatively, it can be cooled to approximately 37°C to 40°C before being transferred to the mold. Before transferring the liquid stearin to the mold, the mold can be coated (e.g., sprayed) with a release agent (such as mineral oil, or another pharmaceutically acceptable release agent). This helps the suppository to be released from the mold once it has cooled / hardened in the mold. Before releasing the hardened suppository from the mold, the suppository can be cooled or chilled to approximately 10°C to 25°C.

[0373] Figure 2 An example of a method for manufacturing rectal suppositories is shown.

[0374] After manufacturing the rectal suppository, it can be stored at a temperature below the melting point of the rectal suppository, for example, less than or equal to about 25°C, or less than or equal to about 8°C, or between about 0°C and 25°C, or between about 0°C and 8°C, or between about 4°C and 8°C.

[0375] The invention will be described in more detail with reference to the following embodiments, but it should be understood that the invention is not limited thereto.

[0376] Example

[0377] Example 1 - Comparison of stearin

[0378] In vitro dissolution tests can be used to provide an indication of the in vivo behavior of 6-TG diffusion and dissolution from rectal suppositories.

[0379] Dissolution of rectal suppositories containing various stearins (and the same level of surfactant) was tested for comparison. The formulations of the rectal suppositories are shown in Table 3. The 6-thioguanine used in Examples 1A, 1B, 2, and 3 was batch B (see Example 4).

[0380] Table 3: Rectal Suppository Preparations

[0381] Dissolution was measured using the conditions shown in Tables 4 and 5. Dissolution tests were performed on multiple samples (e.g., 3 samples), and the average (mean) dissolution of the samples was calculated.

[0382] Table 4: Dissolution Test Conditions

[0383] Table 5: Ultraviolet-Vis Spectrophotometry

[0384] The dissolution results are shown in Table 6, and in Figure 3 It is shown graphically in the middle.

[0385] Suppocire AM, containing 0.5% surfactant (polysorbate 80), showed the fastest dissolution. Suppocire A, containing 0.5% surfactant (polysorbate 80), showed slower dissolution, but when the amount of surfactant (polysorbate 80) was increased (1.5%), the dissolution rate increased, making it similar to that of Suppocire AM.

[0386] Under the test conditions, Suppocire BM exhibited the slowest dissolution. Note that Suppocire BM has a wider melting range than Suppocire A or AM. Dissolution tests were conducted at 37.0°C (±0.5°C), which may have affected the results.

[0387] For comparison, suppositories made from cocoa butter without surfactants (but with the same level of 6-thiobaccoine) showed almost no dissolution or very slow dissolution under the test conditions.

[0388] Table 6: Average dissolution rate (%) over time

[0389] Example 2 - Comparison of Surfactants

[0390] Dissolution of formulations with different surfactants was tested. The formulations are shown in Table 7. The 6-thioguanine used in Examples 5, 6, 7, and 8 was batch C (see Example 4). Dissolution tests were performed using the conditions shown in Tables 4 and 5.

[0391] Table 7: Rectal Suppository Formulations

[0392] The dissolution results are shown in Table 8, and in Figure 4 It is shown graphically in the middle.

[0393] Table 8: Average dissolution rate (%) over time

[0394] The formulation with 1% SPAN 80 (low HLB, approximately 4) showed the slowest dissolution. Dissolution was incomplete even at 90 minutes with an infinite reading. The formulation with 1% SPAN 20 (HLB approximately 9) was faster, but still relatively slow and incomplete.

[0395] Both polysorbate 80 and Brij S20 have HLB values ​​of approximately 15. Formulations containing both polysorbate 80 and Brij S20 exhibit reasonable dissolution rates. However, the formulation containing polysorbate 80 is the fastest, completing dissolution in approximately 30 to 45 minutes.

[0396] Brij S20 is solid at 37°C. It is believed that the melting point of Brij S20 may affect its ability to mix with other ingredients and its dissolution-promoting effect at 37°C. A 90-minute infinity reading of 102% may indicate some variability in the amount of API in the various suppositories prepared with this formulation (e.g., insufficient homogeneity).

[0397] Example 3 - Amount of suspending agent

[0398] It has been found that, especially during scale-up production, 6-thioguanine can settle to the bottom of the mixing vessel. One way to improve this problem has been found to be by adding a suspending agent.

[0399] The uniformity of 6-thioguanine dosage was tested in formulations with different contents of suspending agents (e.g., silica). The formulations are shown in Table 9. The 6-thioguanine used in Examples 9-15 is batch A (see Example 4).

[0400] Table 9: Rectal Suppository Preparations

[0401] In ten batches of suppositories with the same formulation, the amount of 6-thioguanine (API) was tested using ultra-high performance liquid chromatography (UHPLC) by dissolving each suppository in 0.01 M NaOH and purified water, with a reference solution of 6-thioguanine as a control. The amount of 6-thioguanine in each suppository (mg / unit and % of labeled amount (LC), which is 5 mg in this example), the average value and standard deviation of each batch are shown in Tables 10A and 10B, and are presented in... Figure 5 It is shown graphically in the middle.

[0402] Examples 14 and 15 are 9 kg batches of formulation, both of which were single-pass milled. From the mg / unit, LC% and standard deviation, it can be seen that Example 14 (without suspending agent) had difficulty distributing 6-thioguanine throughout the formulation and was difficult to fill / form into units (suppositories) consistently. Examples 9 through 13 are 100 g batches and were not milled. Example 9 (containing 0.10% silica) improved the standard deviation to 8.7. Higher silica content further improved the standard deviation. The API variability was similar among unmilled suppository batches containing 0.25% to 1% silica. The formulation with the lowest standard deviation (1.6) was Example 15, which contained 1% silica and was milled (homogenized).

[0403] Including suspending agents and / or grinding and / or homogenizing the formulation provides a method to reduce the variability of the amount of 6-thioguanine in suppository batches of the same formulation. Low dose variability between suppository batches is ideal for continuous patient dosing.

[0404] Table 10A: SiO₂ with different levels 2 Comparison of dosage unit uniformity - with an average of 1850 mg / suppository ASM (Advanced Methods for Aging) is corrected for weight and moisture content.

[0405] Table 10B: SiO2 with different levels 2 Comparison of dosage unit uniformity - with an average of 1850 mg / suppository ASM (Advanced Methods for Aging) is corrected for weight and moisture content.

[0406] Example 4 - 6-TG Particle Size

[0407] Particle size distribution in individual API and rectal suppository formulations was measured using a Malvern Mastersizer 3000 (Hydro SM). Samples were dispersed in n-heptane containing 0.1% span 80 pre-saturated with 6-thioguanine (to reduce any dissolution of 6-thioguanine particles from the sample).

[0408] The particle size distribution results are shown in Table 11, and the histogram is shown in Table 11. Figure 6A B, C, D, E, F1 and F2.

[0409] The histogram shows the y-axis as bulk density (%), ranging from 0 to 8 (scaled at 0, 2, 4, 6, and 8), and the x-axis as particle size distribution (µm), ranging from 0.01 to 10000.0 (scaled at 0.01, 0.1, 1.0, 10.0, 100.0, 1000.0, and 10000.0).

[0410] Table 11: Particle Size Distribution (PSD)

[0411] Samples A, B, and C are three different batches of 6-thioguanine (not in suppository formulations). Samples A and B have similar particle size distributions (PSDs), but sample C has a significantly different PSD. Histograms show that each of samples A, B, and C has both a secondary peak and a primary peak. In sample C, the primary peak appears to be a secondary peak found in samples A and B. Therefore, the PSDs of the API may differ.

[0412] Sample D is a placebo formulation (containing silica) without 6-thioguanine (unground). The particles measured are silica. The D50 is approximately 36.5, appearing to lie between the secondary and primary peaks of the 6-thioguanine sample. No histograms were given for the placebo batches without silica or 6-thioguanine (i.e., no particles were detected).

[0413] Sample E is a 5 mg suppository formulation (Example 15, comprising silica and 6-thioguanine batch A), which was milled during manufacturing. Given the presence of silica and 6-thioguanine batch A, the expected peaks should correspond to those of samples A and D; however, the formulation was milled during manufacturing. Comparison of histograms and D10, D50, and D90 values ​​for samples A, D, and E shows that the particle size of silica and / or 6-thioguanine appears to have been reduced by milling.

[0414] Samples F1 and F2 are 20 mg 6-thioguanine suppository formulations (containing silica, BHA, and 6-thioguanine batch C) before and after grinding. Comparing the histograms, D10, D50, and D90 of samples C, D, and F1, peaks of silica (sample D) and 6-thioguanine (sample C) were observed in the histogram of F1, with the peak corresponding to silica (sample D) appearing to be dominant. Comparing F1 and F2, both D50 and D90 decreased. However, the decrease in particle size is likely due to the reduction in silica particle size rather than 6-thioguanine, which already had a smaller particle size before grinding.

[0415] Example 5 - Amount of polysorbate 80

[0416] Dissolution of formulations with different levels of polysorbate 80 was tested. The formulations are shown in Table 12. The 6-thioguanine used in Examples 16-22 is batch C (see Example 4).

[0417] Dissolution tests were performed using the conditions shown in Tables 4 and 5.

[0418] Table 12: Rectal Suppository Preparations

[0419] The dissolution results are shown in Table 12, and in Figure 7 It is shown graphically in the middle.

[0420] Table 12: Average Dissolution Rate (%)

[0421] The formulation containing 0% polysorbate 80 had a dissolution rate of only 13% at 60 minutes.

[0422] As the amount of polysorbate 80 increased, improvements in dissolution rate and / or dissolution completion were observed at 60 minutes; for example, 0.25% polysorbate 80 was slower than 0.5% polysorbate 80, while 1% polysorbate 80 was faster than 0.25% and 0.5% polysorbate. However, after 30 minutes, only minor differences were observed between formulations containing 1%, 1.5%, 2%, 2.5%, and 3% polysorbate 80; the main difference was the initial dissolution rate (e.g., in the first 10 minutes).

[0423] Example 6 - Formulation and Manufacturing Method

[0424] Other examples of rectal suppository formulations are shown in Table 13, each suppository providing 5 mg, 10 mg and 20 mg of 6-thioguanine.

[0425] Table 13: Rectal Suppository Preparations The amount of 6-thioguanine can be adjusted based on its potency relative to the USP reference standard.

[0426] The following methods (for example) can be used for batch sizes of 20 kg: 1. Dispense the stearin into a container (1) (preferably under yellow light).

[0427] 2. Heat the fat in the container (1) to about 57°C with slow stirring until the mixture is clear and homogeneous.

[0428] 3. Dispense the BHA into container (1).

[0429] 4. Keep the temperature of the mixture in vessel (1) at 57°C with slow stirring until BHA dissolves.

[0430] 5. Dispense polysorbate 80 into container (1).

[0431] 6. Maintain the temperature while stirring slowly until the mixture is homogeneous.

[0432] 7. Distribute approximately 1 kg of the mixture from vessel (1) into container (1) and 1 kg into container (2).

[0433] 8. Dispense 6-thioguanine (hemihydrate) into container (1) and mix.

[0434] 9. Distribute the silica into the container (1).

[0435] 10. Maintain the temperature while stirring slowly until the mixture is homogeneous.

[0436] 11. While mixing, distribute the contents of container (1) into vessel (1). Rinse container (1) with the contents of container (2) and incorporate them into vessel (1).

[0437] 12. While stirring, reduce the temperature to approximately 45°C.

[0438] 13. The contents of the vessel (1) are pumped into the vessel (2) through a bead mill (preferably the vessel (2) is set to about 45°C) (preferably a heating belt is used on the pump tube).

[0439] 14. Keep the mixture at approximately 45°C while stirring with a top-mounted stirrer.

[0440] 15. Pour the mixture into the hopper of a suppository filling machine with an open agitator, with empty suppository shells already mounted on the feed reel (e.g., Bonapace BP-10 suppository filling machine, set shell capacity to 2.25 cc, target filling mass to 1.85 g, hopper temperature to 38°C to 40°C).

[0441] 16. Pass the filled suppository shells through a suppository sealing machine (e.g., Bonapace BP-11 suppository sealing machine).

[0442] The melting range of the suppository was measured using a Mettler Toledo automated testing instrument at 1 atmosphere. Melting begins at approximately 33°C to 34°C. Due to the suppository's opacity, the melting endpoint is somewhat unclear, but visual observation shows it has completely melted at 36°C.

[0443] Example 7 - Formulation and Manufacturing Method

[0444] Table 14 lists other examples of rectal suppository formulations, each suppository providing 5 mg and 20 mg of 6-thioguanine, with a reduced total weight per suppository.

[0445] 6-Thioguanine from batch C (see Example 4) was used, with a particle size distribution of D10: 2.9 µm, D50: 9.0 µm, and D90: 151.0 µm. This manufacturing method does not include a grinding step.

[0446] Table 14: Rectal Suppository Preparations The amount of 6-thioguanine can be adjusted based on its potency relative to the USP reference standard.

[0447] Rectal suppositories were prepared using the following method: 1. Weigh out the stearin (Suppocire AM) and place it in a container, then heat it to approximately 57°C.

[0448] 2. While maintaining the temperature at approximately 57°C, add the surfactant (polysorbate 80) to the liquid / molten stearin and stir at low speed using a stirrer or homogenizer. For example, mixing can be done using a homogenizer, or it can be done by mechanical stirring. Examples of low speeds are mechanical mixing at approximately 50 to 200 rpm or homogenization at approximately 1000 rpm, but these can be adjusted depending on the equipment used.

[0449] 3. Mix the surfactant (polysorbate 80) and liquid / molten stearin together for about 5 minutes.

[0450] 4. Maintain the temperature and add the suspending agent (Aerosil 200 pharma) while stirring / homogenizing at low speed.

[0451] 5. Continue stirring / homogenizing to reduce the temperature of the liquid / molten mixture to approximately 45°C.

[0452] 6. While stirring / homogenizing the liquid / molten mixture at 45°C, add 6-thioguanine and disperse it.

[0453] 7. Once 6-thioguanine is dispersed (e.g., no visible aggregates), increase the stirring / homogenization speed to break up the invisible agglomerates without substantially introducing air / bubbles into the mixture. Continue stirring / homogenizing in this manner for approximately 10 to 15 minutes. For example, when mixing is carried out by homogenization, a high speed of 4000 rpm+ can be used, or mechanical stirring can be used for approximately 1000 to 2000 rpm. The speed can be adjusted according to the equipment used.

[0454] 8. Prepare a mold for filling and then set it aside. The mold may be sprayed or coated with a release agent (such as mineral oil) to facilitate suppository release.

[0455] 9. Reduce the stirring speed to low and lower the temperature to 35°C.

[0456] 10. Fill the mold with the liquid / molten mixture. The filling temperature of the liquid / molten mixture is approximately 33°C. Note: If the mixture cools below 30°C during filling, it must not be reheated and should be discarded.

[0457] 11. Cool the suppository to approximately 10°C to 25°C. Then demold the hardened suppository.

[0458] Example 8 - Research Plan

[0459] 1. Purpose and Endpoint

[0460] The primary objective of this study was to evaluate the tolerability, pharmacokinetics, and pharmacodynamics of 6-thioguanine rectal suppositories in patients with ulcerative proctitis who had not responded to other treatments.

[0461] Table 15 lists the endpoints to be evaluated.

[0462] Table 15: Endpoint

[0463] 2. Study Design - Patients receiving thioguanine rectal suppositories

[0464] 2.1. Patient inclusion, exclusion, and informed consent

[0465] This part of the study included seven patients with refractory ulcerative proctitis from Canterbury and Auckland. Informed consent was obtained from all patients, and they were treated with thioguanine suppositories 20 mg (5 patients) or 5 mg (2 patients) once daily for 28 days. Blood samples were collected after the first suppository administration for thioguanine pharmacokinetic studies. On day 29, blood samples were collected for flexible sigmoidoscopy, rectal biopsy specimens were obtained, and blood samples were collected again. Inclusion criteria included a diagnosis of active ulcerative proctitis by colonoscopy or flexible sigmoidoscopy within the two months prior to study enrollment. Patients were given time to consider their decision, and some had questions. Baseline flexible sigmoidoscopy was provided for subjects who had not undergone colonoscopy or flexible sigmoidoscopy in the past two months.

[0466] Patients experiencing adverse events considered relevant to treatment with thioguanine suppositories will be excluded during the trial. Patients experiencing adverse events will be advised to contact their local principal investigator by phone. Possible adverse events include leukopenia, hair loss, and nausea / vomiting.

[0467] Patients must not receive thiopurine medications (azathioprine, 6-mercaptopurine, oral thioguanine) during the study and for at least 4 weeks prior to the study. During the study, they may continue to receive other IBD treatments (5-ASA, prednisone, biologics).

[0468] 2.2. Administration of thioguanine rectal suppositories

[0469] Study participants were provided with 28 × 20 mg or 5 mg of 6-thioguanine suppositories (prepared according to Example 7). The first two patients were randomly selected by computer to receive either the 5 mg or 20 mg dose. One of the subsequent five patients received the 5 mg dose, randomly selected by computer, while the remaining patients received the 20 mg dose. Participants were instructed to insert the suppository rectally every night for 28 days.

[0470] 2.3. Sample Collection Date

[0471] Day 1 - The patient will visit the Southern Endoscopy Centre (Christchurch, NZ) and ShoreSurgery (Auckland, NZ). An indwelling intravenous catheter will be placed in the arm or back of the hand. A blood sample will be collected as a baseline pharmacokinetic sample before the first suppository administration. After the first suppository administration, blood samples will be collected at 30 minutes, 1, 2, 4, and 6 hours post-suppository administration for analysis of plasma TG and red blood cell (RBC) 6TGN.

[0472] On day 28 (24 hours before the flexible sigmoidoscopy on day 29), the patient will be given a thioguanine suppository. On the morning of day 29, the subject will travel to the Southern Endoscopy Centre (SEC) (Christchurch, NZ), or Shakespeare Gastroenterology (SG), Shore Surgery (Auckland, NZ). Upon arrival at the SEC or SG, the patient will receive a Fleet enema (laxative enema) to empty the rectum before the endoscopy. An indwelling intravenous catheter will be placed in the arm or back of the hand to collect baseline blood samples for analysis of TG concentration and 6TGN levels in red blood cells (RBCs). The patient will then be transferred to the endoscopy room.

[0473] During a flexible sigmoidoscopy, the rectum is thoroughly flushed with saline solution via the endoscope, and any residual fecal material and any Fleet enema fluid are aspirated to remove it. Once the rectum has been thoroughly flushed and aspirated, 12 small biopsy specimens are taken from the entire rectum and placed in a polypropylene container containing saline and DTT, as described in the assay method below. The polypropylene container is then immediately placed in liquid nitrogen for rapid freezing and transferred to a -80°C freezer for subsequent analysis.

[0474] After a flexible sigmoidoscopy, a 20 mg or 5 mg thioguanine suppository is administered rectally. Blood samples are collected at 30 minutes, 1, 2, 4, and 6 hours after suppository administration for analysis of plasma triglycerides (TG) and red blood cell (RBC) 6TGN.

[0475] 2.4. Validity, safety signals, and compliance

[0476] To identify any initial signals of efficacy, each patient's SCCAI (Simple Clinical Colitis Activity Index) will be assessed on day 1 (baseline), day 8, day 15, and day 29. Visits on days 8 and 15 will be conducted in person by the investigator or research nurse or via telemedicine appointment, including SCCAI scoring and safety checks. Fecal calprotectin levels will be measured on days 1 and 28 as a secondary biomarker of inflammation. At baseline and day 29, endoscopic scores using the Mayo Cognitive Assessment (MCA) and UCEIS scores will be used to grade the severity of proctitis. Any adverse events occurring during treatment will be recorded at the time of SCCAI recording, or at any other time between day 0 and day 29. Specifically, patients will be asked about any new gastrointestinal symptoms, including worsening rectal discomfort or any other new symptoms.

[0477] When administered orally at doses similar to those used in this study, possible side effects of 6-thioguanine include abnormal liver function tests, decreased blood cell counts, nausea and / or vomiting, rash, and infection. Adverse reactions were monitored via weekly questionnaires and weekly blood tests.

[0478] Patients were required to keep a study drug administration diary and return all unused study drugs on day 29.

[0479] 2.5. Blood Sampling

[0480] On day 1 and on day 29 (the day of flexible sigmoidoscopy), an IV indwelling catheter was inserted upon arrival at the SEC or SG, and 5 ml of blood was collected in an EDTA tube before suppository administration. Additional 5 ml blood samples were collected at 30 minutes, 1, 2, 4, and 6 hours after suppository administration. Plasma and RBCs were separated by centrifugation and stored at -80°C for subsequent determination of thioguanine and 6-thioguanine nucleotides.

[0481] 2.6 Milk sampling

[0482] For the relevant patients, breast milk will be collected from both breasts on the day of the flexible sigmoidoscopy (day 29). Breast milk will be collected before and 2, 6, and 9 hours after suppository administration. The breast milk will be transported at 4°C for subsequent determination of thioguanine and 6-thioguanine nucleotides.

[0483] 3. Sample storage

[0484] Store the samples in a -80°C freezer.

[0485] 4.6-Thioguanine and metabolite determination

[0486] Please refer to subsequent chapters of the LC-MS / MS method.

[0487] 5. Summary: Research Center Visit Schedule: The research center's visit schedule is summarized in Table 16.

[0488] Table 16: Research Center Visit Schedule

[0489] 6. Control patients

[0490] In patients receiving oral thiopurine therapy for inflammatory bowel disease, the concentrations of thiopurine and its metabolites in the rectal or colonic tissue are unknown, thus lacking baseline data for comparison with the aforementioned findings. Therefore, tissue drug and metabolite concentrations from control patients are needed.

[0491] 6.1. Patient inclusion, exclusion, and informed consent

[0492] This part of the study included eight patients from Canterbury and Auckland (New Zealand) who underwent routine diagnostic colonoscopies. Four patients received oral 20 mg of thioguanine daily for at least four weeks. Two patients received oral azathioprine for at least four weeks. Two patients had not received mercaptopurine treatment for at least four weeks. Patients were aged 18 to 65 years and were either male or female.

[0493] After obtaining informed consent from the patient, four sets of biopsy specimens were collected from the rectum and colon, except for any other biopsies required for diagnostic purposes.

[0494] During a colonoscopy, the biopsy sites in the rectum and colon are flushed with saline solution and aspirated to remove any residual fecal material. A total of 12 small biopsy specimens are collected and placed in polypropylene containers containing saline and DTT, as described in the assay method below. The polypropylene containers are then immediately placed in liquid nitrogen for rapid freezing and transferred to a -80°C freezer for subsequent analysis.

[0495] 7. LC-MS / MS Analysis Method for 6-TG in Plasma, Red Blood Cells (RBCs) and Colon Tissue - (Mei Zhang, 22-09-2021)

[0496] The patient's whole blood and colon biopsy tissue samples were sent to the Clinical Pharmacology Laboratory (Canterbury Health Laboratory Toxicology Division) of the Faculty of Medicine at the University of Otago (Christchurch), New Zealand, to analyze the 6-TG concentration in plasma, RBCs, and colon tissue.

[0497] 7.1 Sample Collection

[0498] 7.1.1. Whole blood in EDTA tubes for plasma and RBC samples

[0499] 1. This method uses 0.2 mL of plasma and 0.2 mL of compressed RBC to determine 6-TG in plasma and RBC, respectively. To ensure repeatability of analysis if necessary, 5.0 mL of whole blood (EDTA tube) is collected.

[0500] 2. Plasma and RBCs were separated by centrifugation.

[0501] 3. Transfer the plasma to a new polypropylene tube and store the plasma sample at -80°C until it is transported to the laboratory for 6-TG concentration analysis.

[0502] 4. Wash the separated RBCs twice with twice the volume of saline and centrifuge at 1250 g for 5 minutes.

[0503] 5. Count the number of RBCs after washing.

[0504] 6. Transfer the washed RBCs to new polypropylene tubes and store the washed RBC samples at -80°C until transported to the laboratory for 6-TG concentration analysis.

[0505] 7. Plasma and RBC samples were transported to the Department of Toxicology (NZ) of Canterbury Health Laboratories on dry ice to maintain freezing.

[0506] 7.1.2. Colon biopsy tissue

[0507] 1. Collect the colon biopsy tissue into a polypropylene tube.

[0508] 2. Rinse the tissue sample in physiological saline to remove foreign blood and fluid.

[0509] 3. Store tissue samples at -80°C until transported to the laboratory for 6-TG concentration analysis.

[0510] 4. Keep the sample frozen on dry ice and transport it to the Department of Toxicology (NZ) of Canterbury Health Laboratory.

[0511] 7.2 API 4000 LC-MS / MS System and Analysis Conditions

[0512] The Shimadzu LC-20AD HPLC system (shimadzucorporation, Kyoto, Japan) was coupled with an API 4000™ triple quadrupole mass spectrometer (Applied Biosystems, FosterCity, Canada) equipped with a TurboIonSpray® source.

[0513] 6-TG and its isotope-labeled internal standard 6-TG- 13 C2, 15 The ion conversion, declustering potential (DP), collision energy (CE), and collision cell exit potential (CXP) of the N precursor-to-product were optimized.

[0514] Table 17:

[0515] The MS operates in positive ion mode, with flow rates of curtain gas, gas 1, and gas 2 at 20 psi, 45 psi, and 60 psi, respectively. The ion spray voltage is 5000 V, and the source temperature is 500 °C.

[0516] Analyst software (Applied Biosystems, Foster City, Canada) was used to control equipment, coordinate data acquisition, and perform data analysis.

[0517] Analytical column: Phenomenex Luna C18(2) (50 x 2.0 mm ID; 5 μm) (Phenomenex, Torrance, CA, USA).

[0518] Mobile phase: solvent A (0.05% formic acid) and solvent B (methanol), gradient elution for 6 min, flow rate 0.3 mL / min, separating 6-TG and its isotope-labeled internal standard 6-TG on the analytical column. 13 C2,15 N. Gradient elution is shown in Table 18.

[0519] Table 18: Gradient Elution Table

[0520] 7.2.1 6-TG Chromatogram

[0521] Figure 8 The chromatogram of 6-TG is shown.

[0522] 7.3. Materials

[0523] 6-TG, dimethyl sulfoxide (DMSO), DL-dithiothreitol (DTT), sodium chloride, and ammonium bicarbonate were purchased from Sigma Chemical Company (St. Louis, MO, USA).

[0524] Isotope-labeled internal standard, 6-thioguanine- 13 C2, 15 N (6-TG- 13 C2, 15 N) Purchased from Toronto Research Chemicals (Toronto, Canada)

[0525] 70% perchloric acid, HPLC-grade acetonitrile, methanol and formic acid were purchased from British Drug Houses (Poole, UK).

[0526] The phosphate-buffered saline (Dulbecco A) with pH 7.3 was purchased from Oxoid (Basingstoke, UK).

[0527] Distilled deionized water is produced using the Milli-Q Reagent Water System (Millipore, MA, USA).

[0528] Drug-free human plasma and RBCs used for blank assays and for preparing standard curves and quality controls (QC) were obtained from New Zealand Blood Services (Christchurch, New Zealand).

[0529] Drug-free colon tissue used for blank assays and for preparing standard curves and quality controls (QC) was obtained from the Department of Gastroenterology at Christchurch Hospital (Christchurch, New Zealand).

[0530] 7.4. Solution Preparation

[0531] 10 mg / 0.9 mL DL-Dithiothreitol (DTT) aqueous solution - Dissolve 11.1 g DTT in 1000 mL distilled water in a volumetric flask.

[0532] Brine (0.9% sodium chloride aqueous solution) - Dissolve 9.0 g of sodium chloride in 1000 mL of distilled water in a volumetric flask. Store at 4°C.

[0533] 300 mM ammonium bicarbonate (NH4HCO3, ABC) aqueous solution - Dissolve 23.73 g of ABC in 1.0 L of aqueous solution. Store at 4°C.

[0534] 0.05% Formic Acid Aqueous Solution - Dilute 500 µL of formic acid (BDH, 98 / 100%) in 1000 mL of distilled water in a volumetric flask.

[0535] Washed drug-free red blood cells (RBCs): 1. Collect drug-free RBCs from New Zealand Blood Services.

[0536] 2. Wash RBCs in twice the volume of physiological saline and centrifuge at 1250 g for 5 min.

[0537] 3. Count the washed RBCs and adjust the RBC concentration to 8 x 10⁻⁶ using saline. 8 Red blood cells / 0.1 mL RBC solution.

[0538] 4. Store the blank washed RBC solution at -80℃ for later use.

[0539] Drug-free colon tissue homogenate: 1. Collect blank colon tissue from the Department of Gastroenterology at Christchurch Hospital (NZ). Weigh the blank colon tissue.

[0540] 2. Add four times (w / w) of cold PBS and homogenize using Omni Tissue Homogenizer (Omni International, Kennesaw GA, USA).

[0541] 3. Store the blank colon tissue homogenate at -80℃ for later use.

[0542] 7.5. Preparation of Standard Solutions

[0543] 6-TG Stock Solution (0.5 mg / mL) – Dissolve 10 mg of 6-TG in 20 mL of a 1:1 mixture of methanol and dimethyl sulfoxide (DMSO) in a volumetric flask. Two sets of 6-TG stock solutions were prepared, one for the standard curve and one for quality control (QC) samples. All stock solutions were stored at -80°C.

[0544] 6-TG Intermediate Standard Solution (0.1 mg / mL Aqueous Solution) - Dilute 200 µL of 0.5 mg / mL 6-TG stock solution to 1.0 mL with distilled water. Store at -80°C.

[0545] 6-TG- 13 C2, 15 N (internal standard) stock solution (1.0 mg / mL) - to 1.0 mg 6-TG- 13 C2, 15 Add 1.0 mL of a 1:1 mixture of methanol and DMSO to a vial containing N. Store at -80°C.

[0546] Internal standard working solution (1.0 μg / mL, 0.1 M HCl) - Add 50 µL of 1.0 mg / mL 6-TG- 13 C2, 15 Add N stock solution to 50 mL of 0.1 M HCl. Store at -80°C.

[0547] 7.6. Preparation of standard curves and QC for plasma sample analysis

[0548] Standard curve (ST) for 6-TG in plasma: 1. ST6 (400 ng / 0.1 mL): Dilute 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) in a volumetric flask with drug-free plasma to a final volume of 5.0 mL. Store in 100 μL aliquots at -80°C.

[0549] 2. ST5 (200 ng / 0.1 mL): Mix 1.0 mL of ST 6 with 1.0 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0550] 3. ST4 (100 ng / 0.1 mL): Mix 1.0 mL of ST 5 with 1.0 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0551] 4. ST3 (50 ng / 0.1 mL): Mix 1.0 mL of ST4 with 1.0 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0552] 5. ST2 (25 ng / 0.1 mL): Mix 1.0 mL of ST3 with 1.0 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0553] 6. ST1 (10 ng / 0.1 mL): Mix 1.0 mL of ST2 with 1.25 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0554] QC for 6-TG in plasma: 1. QC 3 (400 ng / 0.1 mL): Dilute 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) in a volumetric flask with drug-free plasma to a final volume of 5.0 mL. Store in 100 μL aliquots at -80°C.

[0555] 2. QC 2 (100 ng / 0.1 mL): Mix 1.0 mL of QC 3 with 3.0 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0556] 3. QC 1 (10 ng / 0.1 mL): Mix 0.5 mL of QC 2 with 4.5 mL of drug-free plasma. Store in 100 μL aliquots at -80°C.

[0557] 7.7. Preparation of Standard Curves and QC for RBC Sample Analysis

[0558] Standard curve (ST) for 6-TG in RBC: 1.ST6 (400 ng / 8x10 8 RBC / 0.1 mL: Add 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) to a volumetric flask, and then add 8 x 10 mL of washed drug-free RBC solution. 8 Dilute RBC (0.1 mL RBC solution) to 5.0 mL. Store in 100 μL aliquots at -80 °C.

[0559] 2.ST5 (200 ng / 8x10 8 RBC / 0.1 mL): Mix 1.0 mL of ST 6 with 1.0 mL of washed, drug-free RBC solution. Store in 100 μL aliquots at -80 °C.

[0560] 3. ST4 (100 ng / 8x10 8RBC / 0.1 mL): Mix 1.0 mL of ST 5 with 1.0 mL of washed, drug-free RBC solution. Store in 100 μL aliquots at -80 °C.

[0561] 4.ST3 (50 ng / 8x10 8 RBC / 0.1 mL): Mix 1.0 mL of ST 4 with 1.0 mL of washed, drug-free RBC solution. Store in 100 μL aliquots at -80°C.

[0562] 5.ST2 (25 ng / 8x10 8 RBC / 0.1 mL): Mix 1.0 mL of ST 3 with 1.0 mL of washed, drug-free RBC solution. Store in 100 μL aliquots at -80 °C.

[0563] 6.ST1 (10 ng / 8x10 8 RBC / 0.1 mL): Mix 1.0 mL ST 2 with 1.25 mL of washed drug-free RBC solution. Store in 100 μL aliquots at -80 °C.

[0564] QC in 6-TG of RBC:

[0565] 1. QC 3 (400 ng / 8x10 8 RBC / 0.1 mL: Add 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) to a volumetric flask, and then add 8 x 10 mL of washed drug-free RBC solution. 8 Dilute RBC (0.1 mL RBC solution) to 5.0 mL. Store in 100 μL aliquots at -80 °C.

[0566] 2. QC 2 (100 ng / 8x10 8 RBC / 0.1 mL): Mix 1.0 mL of QC 3 with 3.0 mL of washed drug-free RBC solution. Store in 100 μL aliquots at -80°C.

[0567] 3. QC 1 (10 ng / 8x10 8 RBC / 0.1 mL): Mix 0.5 mL of QC 2 with 4.5 mL of washed, drug-free RBC solution. Store in 100 μL aliquots at -80°C.

[0568] 7.8. Preparation of standard curves and QC for colon tissue sample analysis

[0569] Standard curve (ST) of 6-TG in colon tissue

[0570] 1. ST6 (400 ng / 0.1 mL): Dilute 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) in a volumetric flask with blank colon tissue homogenate to a final volume of 5.0 mL. Store in 100 μL aliquots at -80°C.

[0571] 2. ST5 (200 ng / 0.1 mL): Mix 1.0 mL of ST 6 with 1.0 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0572] 3. ST4 (100 ng / 0.1 mL): Mix 1.0 mL of ST 5 with 1.0 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0573] 4. ST3 (50 ng / 0.1 mL): Mix 1.0 mL of ST4 with 1.0 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0574] 5. ST2 (25 ng / 0.1 mL): Mix 1.0 mL of ST3 with 1.0 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0575] 6. ST1 (10 ng / 0.1 mL): Mix 1.0 mL of ST2 with 1.25 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0576] QC of 6-TG in colon tissue

[0577] 1. QC 3 (400 ng / 0.1 mL): Dilute 200 µL of 6-TG intermediate standard solution (0.1 mg / mL 6-Tg aqueous solution) in a volumetric flask with blank colon tissue homogenate to a final volume of 5.0 mL. Store in 100 μL aliquots at -80°C.

[0578] 2. QC 2 (100 ng / 0.1 mL): Mix 1.0 mL of QC 3 with 3.0 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0579] 3. QC 1 (10 ng / 0.1 mL): Mix 0.5 mL of QC 2 with 4.5 mL of blank colon tissue homogenate. Store in 100 μL aliquots at -80°C.

[0580] 7.9. Sample Preparation

[0581] plasma samples

[0582] 1. Place 100 μL of plasma (blank plasma, plasma ST, plasma QC, or patient plasma sample) into a 1.5 mL plastic centrifuge tube.

[0583] 2. Add 50 μL of internal standard working solution (1.0 μg / mL 6-TG- 13 C2, 15 N (soluble in 0.1M HCl), brief vortex

[0584] 3. Add 0.9 mL of 10 mg / 0.9 mL DL-dithiothreitol (DTT).

[0585] 4. Add 100 µL of 70% perchloric acid and immediately vortex for 30 seconds.

[0586] Centrifuge at 5.15000 g for 5 min.

[0587] 6. Mix 50 μL of supernatant with 200 μL of 300 mM ammonium bicarbonate to neutralize the sample.

[0588] 7. Inject 10 μL of the sample into LC-MS / MS.

[0589] RBC and colon tissue samples

[0590] Intracellular 6-thioguanine nucleotide (6-TGN) is the major metabolite of intracellular 6-TG. To avoid underestimating bioavailability, 6-TGN in RBCs and colon tissues is converted to free 6-TG by heated acid hydrolysis before analyzing 6-TG concentration.

[0591] RBC sample

[0592] 1. Centrifuge whole blood to separate RBCs.

[0593] 2. Wash RBCs with twice the volume of saline solution and centrifuge at 1250 g for 5 min.

[0594] 3. Count the washed RBCs and normalize the 6-TG concentration to 8 x 10⁻⁶. 8 RBC.

[0595] 4. Place 100 μL of RBCs (blank RBCs, RBC STs, RBC QCs, or patient RBC samples) into a 1.5 mL plastic centrifuge tube.

[0596] 5. Add 50 μL of internal standard working solution and vortex briefly.

[0597] 6. Add 0.9 mL of 10 mg / 0.9 mL DTT.

[0598] 7. Add 100 µL of 70% perchloric acid and immediately vortex for 30 seconds.

[0599] Centrifuge at 8.15000 g for 5 min.

[0600] 9. Transfer 320 µL to a 96-well plate, cover the plate with an aluminum foil sealant, and heat in a Block Heater at 120 °C for 60 min.

[0601] 10. Immediately and carefully remove the aluminum foil sealant.

[0602] 11. After cooling, mix 50 μL of the supernatant with 200 μL of 300 mM ammonium bicarbonate to neutralize the sample.

[0603] 12. Inject 10 μL of the sample into LC-MS / MS.

[0604] colon tissue sample

[0605] 1. Homogenization of patient tissue samples Weigh the colon biopsy tissue.

[0606] Add four times (w / w) of cold PBS to the colon biopsy tissue sample and homogenize using Omni Tissue Homogenizer.

[0607] 2. Place 100 μL of tissue homogenate (blank tissue homogenate, ST tissue homogenate, QC tissue homogenate, or patient tissue homogenate sample) into a 1.5 mL centrifuge tube.

[0608] 3. Add 50 μL of internal standard working solution and vortex briefly.

[0609] 4. Add 0.9 mL of 10 mg / 0.9 mL DTT.

[0610] 5. Add 100 µL of 70% perchloric acid and immediately vortex for 30 seconds.

[0611] 6. Centrifuge at 15000 g for 5 min.

[0612] 7. Transfer 320 µL of supernatant to a 96-well plate, cover the plate with aluminum foil sealing film, and heat in a Block Heater at 120 °C for 60 min.

[0613] 8. Immediately and carefully remove the aluminum foil sealant.

[0614] 9. After cooling, mix 50 μL of the supernatant with 200 μL of 300 mM ammonium bicarbonate to neutralize the sample.

[0615] 10. Inject 10 μL of the sample into LC-MS / MS.

[0616] result

[0617] The results of the first two patients in this study are shown in Table 19.

[0618] Table 19: Outcomes of Patient 1 and Patient 2

[0619] Ulcerative proctitis is an inflammatory bowel disease that causes inflammation and ulceration in the rectal region. Symptoms include diarrhea, urgency to defecate, and rectal bleeding.

[0620] The UCEIS (Ulcerative Colitis Endoscopic Severity Index) is based on three scoring items, calculated using a simple composite method: vascular texture (0 to 2 points), bleeding (0 to 3 points), and erosion and ulceration (0 to 3 points). The UCEIS score ranges from 0 to 8. The UCEIS score can be categorized as follows: 0 to 1 points: remission; 2 to 4 points: mild disease activity; 5 to 6 points: moderate disease activity; 7 to 8 points: severe disease activity (see, for example, Ikeya et al. J Crohns Colitis. 2016 Mar; 10(3):286-295).

[0621] The Mayo endoscopic score (Mayo ES) ranges from 0 to 3 and can be divided into four categories: 0, normal or inactive disease; 1, mild lesion with erythema, reduced vascularity, and mild fragility; 2, moderate lesion with significant erythema, loss of vascularity, fragility, and erosion; 3, severe lesion with spontaneous bleeding and ulceration (see, for example, Ikeya et al. J Crohns Colitis. Mar 2016; 10(3): 286-295).

[0622] The SCCAI (Simple Clinical Colitis Activity Index) is a questionnaire completed by patients that indicates the severity of colitis symptoms and whether the disease is in an active or remission phase. Questions are based on bowel movement frequency (daytime), bowel movement frequency (nighttime), urgency, rectal bleeding, overall health status, and extraintestinal manifestations (e.g., musculoskeletal disorders such as arthritis, eye disorders such as uveitis and episcleritis, skin disorders such as pyoderma gangrenosum and erythema nodosum, and oral cavity such as aphthous stomatitis). Scores range from 0 to 20.

[0623] A SCCAI score below 2.5 is considered to be in remission (see, for example, Bewtra et al., “An Optimized Patient-reported Ulcerative Colitis Disease Activity Measure Derived from the Mayo Score and the Simple Clinical Colitis Activity Index”, Inflammatory Bowel Dis. 2014 Jun; 20(6): 1070-1078). Clinical response is defined as a decrease in the Simple Clinical Colitis Activity Index (SCCAI) score of 3 or more from baseline (see, for example, Crouwel et al., “Rectally Administrated Thioguanine for Distal Ulcerative Colitis: A Multicenter Case Series”, Inflammatory Bowel Diseases, Vol. 29, No. 6, June 2023, pp. 1000-1004).

[0624] Patient 1 received suppositories containing 20 mg of 6-thioguanine daily. Patient 1 had more severe ulcerative proctitis, with an initial SCCAI score of 10. Patient 1 had a clinical response, showing a decrease of 4 points from baseline in the SCCAI score within 8 days and a decrease of 6 points from baseline within 15 days. The SCCAI score remained at 4 points on day 22, but slightly increased to 5 points on day 29. This was due to the patient's transition from loose stools associated with ulcerative proctitis to constipation, resulting in bloating, discomfort, and straining during defecation, which in turn led to persistent bleeding. These factors caused a false increase in the SCCAI score. A transition to constipation is not uncommon while ulcerative proctitis is being treated. The patient had received separate treatment for constipation. The endoscopic appearance of the rectum showed a very significant improvement, with the Mayo score decreasing from 3 points at the start of the test to 1 point on day 29. The UCEIS score also decreased from 5 points at the start of the test to 2 points on day 29.

[0625] Patient 2 received suppositories containing 5 mg of 6-thioguanine daily. Despite receiving a low dose of 6-thioguanine, the patient had a clinical response according to the SCCAI score and achieved remission within 8 days of starting treatment, maintaining remission on days 15 and 22. Unfortunately, the patient's SCCAI score worsened to 4 on day 29. As with Patient 1, symptoms and severity can change throughout treatment and may be partly due to external factors such as stress and lifestyle. Endoscopic appearance of the rectum showed improvement, and the UCEIS score decreased from 6 at the start of testing to 5 on day 29.

[0626] Table 20: Pharmacokinetics (PK) on Day 1

[0627] Referring to the results in Table 20, the lower limit of quantification (LLOQ) for 6-TGN in red blood cells (RBCs) was 75 pmol / 8. 10^8 RBCs. The LLOQ for serum 6-TG was 2.5 ng / mL. Serum 6-TG concentrations below the LLOQ were obtained by extrapolation from a standard curve at levels below the LLOQ, therefore these values ​​may be subject to significant bias. The analyst was blinded to the patient / drug administration.

[0628] The PK data in Table 20 were obtained from blood samples taken before or after the first administration of the rectal suppository on day 1. Therefore, the results should not be considered as a steady state.

[0629] On day 1, 6-TGN was not detected in either patient / dose group.

[0630] 6-TG was detectable in the serum of patient 1 (20 mg dose), but below the limit of quantitation, so the results may be significantly biased. 6-TG was not detected in patient 2 (5 mg dose), which may be related to the lower dose and / or that the patient's mucosal inflammation may have been less severe than in patient 1, allowing less drug to be directly transported into the bloodstream.

[0631] Although the invention has been described in detail with reference to specific embodiments thereof, it will be apparent to those skilled in the art that various changes and modifications may be made therein without departing from the spirit and scope of the invention.

[0632] Numbered paragraphs: 1. A rectal suppository comprising: 6-Thioguanine, At least one stearin, and At least one surfactant.

[0633] 2. The suppository according to paragraph 1, wherein the suppository contains about 1 mg to 30 mg of 6-thioguanine, or about 3 mg to 25 mg of 6-thioguanine, or about 5 mg to 20 mg of 6-thioguanine.

[0634] 3. A rectal suppository containing approximately 1 mg to 9 mg of 6-thioguanine.

[0635] 4. The suppository according to paragraph 3, wherein the suppository comprises at least one stearin.

[0636] 5. The suppository according to any one of paragraphs 1, 2 or 4 above, wherein the suppository comprises about 80% w / w to 99.9% w / w stearin, or about 85% w / w to 99.5% w / w stearin, or about 90% w / w to 99.5% w / w stearin, or about 95% w / w to 99.5% w / w stearin.

[0637] 6. The suppository according to any one of paragraphs 1, 2, 4 or 5, wherein the stearin comprises less than about 10 mol% of one or more unsaturated fatty acids, or less than about 5 mol% of one or more unsaturated fatty acids, or less than about 2 mol% of one or more unsaturated fatty acids, or less than about 1 mol% of one or more unsaturated fatty acids.

[0638] 7. The suppository according to any one of paragraphs 1, 2 or 4 to 6, wherein the hydroxyl value of the stearin is less than 40 mgKOH / g.

[0639] 8. The suppository according to any one of paragraphs 3 to 7, wherein the suppository comprises at least one surfactant.

[0640] 9. The suppository according to any one of paragraphs 1, 2, or 8, wherein the suppository comprises about 0.1% w / w to 20% w / w of a surfactant, about 0.1% w / w to 15% w / w of a surfactant, or about 0.1% w / w to 10% w / w of a surfactant, or about 0.1% w / w to 5% w / w of a surfactant, or about 0.2% w / w to 3% w / w of a surfactant, or about 0.4% w / w to 3% w / w of a surfactant, or about 0.5% w / w to 3% w / w of a surfactant, or about 0.5% w / w to 2% w / w of a surfactant.

[0641] 10. The suppository according to any one of paragraphs 1, 2, 8 or 9, wherein the hydrophilic-lipophilic balance (HLB) value of the surfactant is equal to or greater than about 10.

[0642] 11. The suppository according to any one of paragraphs 1, 2, or 8 to 10, wherein the surfactant is selected from any one or more of the following: polysorbate, polyoxyethylene derivatives of natural or hydrogenated vegetable oils, alkyl / dialkyl sulfates, sulfonates or sulfosuccinates, polyoxyethylene fatty acid esters, phospholipids, transesterification products of natural vegetable oil triglycerides and polyalkylene polyols, sorbitan fatty acid esters, pentaerythritol fatty acid esters, polyoxyethylene glycol alkyl ethers and / or polyoxyethylene glycol alkyl esters, sucrose esters, ethoxylated fatty alcohols, and fatty acid salts.

[0643] 12. The suppository according to any one of paragraphs 1, 2, or 8 to 11, wherein the surfactant is polysorbate 80.

[0644] 13. The suppository according to any one of paragraphs 1 to 12, wherein the suppository comprises a suspending agent.

[0645] 14. The suppository according to paragraph 13, wherein the suspending agent is present at about 0.05% w / w to 10% w / w, or about 0.1% w / w to 5% w / w, or about 0.1% w / w to 4% w / w, or about 0.1% w / w to 3% w / w, or about 0.1% w / w to 2% w / w, or about 0.1% w / w to 1% w / w.

[0646] 15. The suppository according to paragraph 13 or 14, wherein the suspending agent is selected from one or more of the following: silica, clay (including purified and / or refined clay), aluminum monostearate, and magnesium stearate.

[0647] 16. The suppository according to any one of paragraphs 1 to 15, wherein the suppository comprises an antioxidant.

[0648] 17. The suppository according to any one of paragraphs 1 to 16, wherein the total weight of the suppository is about 800 mg to 3000 mg.

[0649] 18. The suppository according to any one of paragraphs 1 to 17, wherein the total weight of the suppository is about 1,500 mg to 3,000 mg.

[0650] 19. A method for preventing, treating and / or controlling inflammatory bowel disease in a subject, the method comprising administering to a subject in need any of paragraphs 1 to 18 a rectal suppository.

[0651] 20. A method for preventing, treating, and / or controlling inflammatory bowel disease in a subject, the method comprising administering about 1 mg to 9 mg of 6-thioguanine in the form of a suppository to the rectum of the subject.

[0652] 21. Use of any rectal suppository containing 6-thioguanine as described in paragraphs 1 to 18 in the manufacture of a medicament for the prevention, treatment and / or control of inflammatory bowel disease.

[0653] 22. Use of rectal suppositories containing 6-thioguanine in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease, wherein said suppository contains about 1 mg to 9 mg of 6-thioguanine.

[0654] 23. The method or use according to any one of paragraphs 19 to 22, wherein the inflammatory bowel disease involves the rectum.

[0655] 24. The method or use according to any one of paragraphs 19 to 23, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease involving the rectum.

[0656] 25. The method or use according to paragraph 24, wherein the ulcerative colitis is ulcerative proctitis, rectosigmoid colitis, and / or left colitis.

[0657] 26. A method for manufacturing a rectal suppository, the method comprising the following steps: (a) Heating stearin to turn it into liquid stearin. (b) Add at least one surfactant to the liquid stearin. (c) Add 6-thioguanine to the liquid stearin. (d) Form the stearin into a rectal suppository. Step (b) may be performed before or after step (c).

Claims

1. A rectal suppository comprising: 6-Thioguanine, At least one stearin, and At least one surfactant.

2. The suppository according to claim 1, wherein, The suppository contains approximately 1 mg to 30 mg of 6-thioguanine.

3. A rectal suppository containing approximately 1 mg to 9 mg of 6-thioguanine.

4. The suppository according to claim 3, wherein, The suppository contains at least one type of stearin.

5. The suppository according to any one of claims 1, 2, or 4, wherein, The suppository contains approximately 80% w / w to 99.9% w / w stearin.

6. The suppository according to any one of claims 1, 2, 4 or 5, wherein, The stearin contains less than about 10 mol% of one or more unsaturated fatty acids.

7. The suppository according to any one of claims 1, 2, or 4 to 6, wherein, The stearin has a hydroxyl value of less than 40 mgKOH / g.

8. The suppository according to any one of claims 3 to 7, wherein, The suppository contains at least one surfactant.

9. The suppository according to any one of claims 1, 2, or 8, wherein, The suppository contains approximately 0.1% w / w to 20% w / w of surfactant.

10. The suppository according to any one of claims 1, 2, 8 or 9, wherein, The surfactant has a hydrophilic-lipophilic balance (HLB) value equal to or greater than about 10.

11. The suppository according to any one of claims 1, 2, or 8 to 10, wherein, The surfactant is selected from any one or more of the following groups: polysorbate, polyoxyethylene derivatives of natural or hydrogenated vegetable oils, alkyl / dialkyl sulfates, sulfonates or sulfosuccinates, polyoxyethylene fatty acid esters, phospholipids, transesterification products of natural vegetable oil triglycerides and polyalkylene polyols, sorbitan fatty acid esters, pentaerythritol fatty acid esters, polyoxyethylene glycol alkyl ethers and / or polyoxyethylene glycol alkyl esters, sucrose esters, ethoxylated fatty alcohols, and fatty acid salts.

12. The suppository according to any one of claims 1, 2, or 8 to 11, wherein, The surfactant is polysorbate 80.

13. The suppository according to any one of claims 1 to 12, wherein, The suppository contains a suspending agent.

14. The suppository according to any one of claims 1 to 13, wherein, The suspending agent is present at a concentration of approximately 0.05% w / w to 10% w / w.

15. The suppository according to claim 13 or 14, wherein, The suspending agent is selected from one or more of the following: silica, clay, aluminum monostearate, and magnesium stearate.

16. The suppository according to any one of claims 1 to 15, wherein, The suppository contains an antioxidant.

17. The suppository according to any one of claims 1 to 16, wherein, The total weight of the suppositories is approximately 800 mg to 3000 mg.

18. The suppository according to any one of claims 1 to 17, wherein, The total weight of the suppository is approximately 1500 mg to 3000 mg.

19. A method for preventing, treating and / or controlling inflammatory bowel disease in a subject, the method comprising administering to a subject in need the rectal suppository of any one of claims 1 to 18.

20. A method for preventing, treating, and / or controlling inflammatory bowel disease in a subject, the method comprising administering about 1 mg to 9 mg of 6-thioguanine in the form of a suppository to the rectum of the subject.

21. Use of the rectal suppository containing 6-thioguanine according to any one of claims 1 to 18 in the manufacture of a medicament for the prevention, treatment and / or control of inflammatory bowel disease.

22. Use of rectal suppositories containing 6-thioguanine in the manufacture of medicaments for the prevention, treatment and / or control of inflammatory bowel disease, wherein said suppository contains about 1 mg to 9 mg of 6-thioguanine.

23. The method or use according to any one of claims 19 to 22, wherein, The inflammatory bowel disease involves the rectum.

24. The method or use according to any one of claims 19 to 23, wherein, The inflammatory bowel disease referred to is ulcerative colitis or Crohn's disease that affects the rectum.

25. The method or use according to claim 24, wherein, The ulcerative colitis refers to ulcerative proctitis, rectosigmoid colitis, and / or left colitis.

26. A method for manufacturing a rectal suppository, the method comprising the following steps: (a) Heating stearin to turn it into liquid stearin. (b) Add at least one surfactant to the liquid stearin. (c) Add 6-thioguanine to the liquid stearin. (d) Form the stearin into a rectal suppository. Step (b) may be performed before or after step (c).

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