Polymorphic forms of deoxycytidine, compositions comprising the same and uses

Abstract

To provide deoxycytidine compositions with reduced residual solvent content.SOLUTION: Provided is a composition comprising Form B deoxycytidine and at least one residual solvent, the form B deoxycytidine being characterized by a powder X-ray diffraction pattern comprising peaks at 13.7°, 17.2°, 18.0°, 19.2° and 22.8 (2θ) (± 0.2°(2θ)), the at least one residual solvent being selected from the group consisting of methanol, toluene, methylene chloride, ethanol, tert-butylmethyl ether, acetone, ethyl acetate, and n-heptane, and the composition being stable for at least 3 months at 25±2°C and 60±5% relative humidity.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Patent Application No. 62 / 888,893, filed August 19, 2019. No. 60 / 019,493, filed on Dec. 1, 2003, which is incorporated herein in its entirety.

[0002] The present invention relates to polymorphic forms of deoxycytidine; polymorphic forms of deoxycytidine and ICH limits at least one residual solvent at a concentration of less than 10% of the original concentration; and Diseases or disorders characterized by nucleotide pools, particularly mitochondrial DNA depletion syndromes polymorphic forms of deoxycytidine and less than 10% of the ICH limit The present invention relates to a method of treating a subject with a composition comprising at least one residual solvent at a concentration of [Background technology]

[0003] Polymorphs are two substances that have different arrangements and / or different conformations of the molecules within the crystal lattice. If the solvent molecules are contained within the crystal lattice, the resulting crystals will be They are called pseudopolymorphs or solvates. If the solvent molecules in the crystal structure are water molecules, then they are pseudopolymorphs. Polymorphs and pseudopolymorphs are solids that result from packing, and various thermodynamic, spectroscopic, interfacial and mechanical properties (see Non-Patent Document 1 They exhibit different physical properties, including: Polymorphic and pseudopolymorphic forms of benzodiazepine (presumably benzodiazepine) may be present when administered alone or in formulations (most commonly Formulated as a final or finished dosage form, also known as a pharmaceutical composition It is well known when pharmaceutical formulations are used, for example, to evaluate the solubility, stability, flowability, friability and This can affect the compressibility and safety and efficacy of the formulation (see, for example, Non-Patent Document 2). See the reference.

[0004] Deoxycytidine (2´-deoxycytidine, dC) is a nucleotide analogue of deoxythymidine (dT). When used in combination, they are effective in treating certain mitochondrial DNA depletion syndromes. Mitochondrial DNA depletion syndrome (MDS), a subgroup of mitochondrial diseases, is Mitochondrial DNA (MDS) is a condition characterized by a decrease in mitochondrial DNA (mtDNA) copy number in tissues and the A frequent cause of severe childhood encephalomyopathies characterized by insufficient synthesis of the endothelial RC complex Yes (Hirano, et al. 2001). TK2, DGUOK, POLG, POL G2, SCLA25A4, MPV17, RRM2B, SUCLA2, SUCLG1, TY Mutations in several nuclear genes, including MP, OPA1, and ClOorfl (PEOl) has been identified as a cause of infantile MDS (Bourdon, et al. 2007; Copeland2008;Elpeleg,et al.2005;Mandel,e t al.2001;Naviaux and Nguyen2004;Osterga ard,et al.2007;Saada,et al.2003;Sarzi,et al.2007; Spinazzola, et al., 2006). Furthermore, these Mutations in nuclear genes also occur in multiple mtDNA regions with or without mtDNA depletion. can cause deletion of l.2012;Longley,et al.2006;Nishino,et al. 1999;Paradas,et al.2012;Ronchi,et al.201 2;Spelbrink,et al.2001;Tyynismaa,et al.2 009;Tyynismaa,et al.2012;Van Goethem,et al. 2001).

[0005] One of these genes is TK2, which encodes thymidine kinase (TK2), i.e. , deoxythymidine monophosphate (dTMP) and deoxycytidine monophosphate (dCMP) Phosphate pyrimidine nucleosides (thymidine and deoxycytidine) to produce It encodes a mitochondrial enzyme required for mitochondrial synthesis (Saada, et al. 2001). The mutation in K2 disrupts the deoxyribonuclease, a component for mRNA replication and repair. Mitochondrial nucleoside / nucleoside required for nucleotide triphosphate (dNTP) synthesis Impairs the otide salvage pathway.

[0006] Patients diagnosed with TK2 deficiency are treated with a combination of deoxycytidine and deoxythymidine. The dose administered is relatively high, for example, 400 mg / kg / day of each of oxycytidine and deoxythymidine Therefore, a 100 kg patient should receive a maximum of 80 g of active substance per day (40 g of deodorant). 40 g of oxycytidine and 40 g of deoxythymidine) can be administered. The need to minimize the amount of residual solvent resulting from the manufacture of the active ingredient is important. are classified as ICH Class 1 (i.e., solvents to be avoided), Class 2 (i.e., solvents to be restricted), These include Class 1 (i.e., solvents with low toxicity) and Class 2 (i.e., solvents with low toxicity). The classification of solvents by chemical structure is known. The ICH guidelines define Class 2 and Class 3 residues. The permissible daily intake (PDE) and concentration limits for solvents are specified. 1 solvents unless unavoidable, and the ICH guidelines recommend that concentrations of these solvents be very low. should not be used in manufacturing unless limited to extremely low levels.

[0007] Therefore, residual There is a need for improved manufacturing processes that provide deoxycytidine compositions with reduced solvent content. It is considered essential. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] WO2016205671 [Non-patent literature]

[0009] [Non-Patent Document 1] H. Brittain, Polymorphism in Pharmaceutical Solids, Marcel Dekker, New York, NY, 1999, pp.1-2 [Non-patent document 2] Knapman,K Modem Drug Discovery,Mar.2000:53 Summary of the Invention

[0010] In one embodiment, the present invention provides deoxycytidine in form B. Cytidine has the X-ray powder diffraction (XRPD) pattern shown in Figure 2. In one embodiment, Form B of deoxycytidine has the following positions: 13.7°, 17.2°, 18.0°, 19.2°, and and 22.8° (2θ) (±0.2° (2θ)). In one embodiment, the XRPD pattern of Form B is 11.5°, 11.8°, 13 .7°, 17.2°, 18.0°, 19.2°, 20.2°, 21.1°, 21.4°, One or more peaks at 21.8° and 22.8° (2θ) (±0.2° (2θ)) Further includes:

[0011] Deoxycytidine Form B of the present invention can be prepared by a large-scale manufacturing process. and contains minimal residual solvent. In one embodiment, deoxycytidine in Form B is about Less than 100 ppm ethyl acetate and / or less than about 100 ppm n-heptane and and / or has less than about 200 ppm ethanol.

[0012] Deoxycytidine Form B of the present invention may, for example, remain at least 100% when stored in a sealed container at room temperature. Both are stable for approximately six months. Stability is confirmed by several reactions, including the conversion of Form A to deoxycytidine. It can be measured by the method.

[0013] Form B deoxycytidine can be substantially pure, i.e., about 95% by weight or greater. In another embodiment, the mixture may comprise a form of deoxycytidine having form B. Thus, isolated Form B is provided.

[0014] In another aspect, the present invention is a method for preparing deoxycytidine form B. (a) providing crude deoxycytidine; (b) subjecting the crude deoxycytidine to ethanol; (c) contacting the first mixture with ethanol to form a first mixture; and (d) contacting the residue with purified water to form a second distillate. (e) increasing the internal temperature of the second mixture to at least about 40° C. (f) heating the ethanol, ethyl acetate, and heptane to a second temperature; (g) adding the mixture to form a third mixture; and (g) cooling the third mixture to form a (h) forming crystals of deoxycytidine in form B; and separating the crystals from the third mixture.

[0015] The methods of manufacture described herein may be used in combination with, for example, commercial chemical suppliers or previous manufacturing methods. In comparison, the present method provides a deoxycytidine composition with less residual solvent. Approximately 10% of the Class 2 and Class 3 residual solvent concentrations allowed by CH guidelines The method is also optimized to provide a deoxycytidine composition having less than 1000 kJ / ml. , according to validated methods, undetectable levels of certain Class 1 solvents (e.g., 1,2- Deoxycytidine compositions containing methyl methyl ether (dichloroethane) are provided.

[0016] In one embodiment, the composition comprises deoxycytidine form B and ICH class 2 and class 3. and at least one residual solvent selected from the solvents of Class 3, The solvent concentration is less than about 10% of the ICH concentration limit for each residual solvent. Preferred Class 2 and Class 3 residual solvents include methanol, toluene, methylene chloride, ethanol, t ert-butyl methyl ether (TBME), acetone, ethyl acetate and n-heptane Includes:

[0017] In a specific embodiment, the composition comprises deoxycytidine Form B and (a) 1 ppm (b) toluene from 1 ppm to about 89 ppm; (c) toluene from 1 ppm to about 300 ppm; pm to about 600 ppm methylene chloride, (d) 1 ppm to about 500 ppm ethanol, (e) TBME from 1 ppm to approximately 500 ppm, (f) acetone from 1 ppm to approximately 500 ppm (g) 1 ppm to about 500 ppm of ethyl acetate, and (h) 1 ppm to about 500 ppm of pm n-heptane, and

[0018] In another specific embodiment, the composition comprises deoxycytidine form B and 300 pp Methanol less than 89 ppm, toluene less than 60 ppm, methylene chloride less than 50 ppm Ethanol less than 0 ppm, TBME less than 500 ppm, Acetone less than 500 ppm , 500 ppm or less of ethyl acetate and 500 ppm or less of n-heptane.

[0019] In a preferred embodiment, the composition contains undetectable levels of Class 1 solvents, particularly Class 1, It has 2-dichloroethane.

[0020] In yet another embodiment, a substantially pure Form B deoxycytidine and at least A pharmaceutical composition is provided that includes a pharmaceutically acceptable carrier. It may further comprise thymidine.

[0021] In one embodiment, the pharmaceutical composition comprises deoxycytidine Form B and at least one a residual solvent and at least one pharmaceutically acceptable carrier, The distillation solvents are selected from ICH Class 2 and Class 3 solvents and contain at least one residual solvent. The concentration of the solvent is less than approximately 10% of the ICH concentration limit for each residual solvent.

[0022] In a specific embodiment, the pharmaceutical composition comprises deoxycytidine Form B and at least a pharmaceutically acceptable carrier; (a) 1 ppm to about 300 ppm of methanol; (b) ) 1 ppm to about 89 ppm of toluene, (c) 1 ppm to about 60 ppm of methylene chloride, (d) 1 ppm to approximately 500 ppm of ethanol, (e) 1 ppm to approximately 500 ppm of TB ME, (f) 1 ppm to about 500 ppm of acetone, (g) 1 ppm to about 500 ppm of ethyl acetate, and (h) 1 ppm to about 500 ppm of n-heptane Includes one and.

[0023] In another specific embodiment, the pharmaceutical composition contains no more than 300 ppm of methanol, ppm or less of toluene, 60 ppm or less of methylene chloride, 500 ppm or less of ethanol , TBME less than 500 ppm, acetone less than 500 ppm, acetic acid less than 500 ppm Contains ethyl and up to 500 ppm of n-heptane.

[0024] In a preferred embodiment, the pharmaceutical composition contains undetectable levels of Class 1 solvents, particularly It has 1,2-dichloroethane.

[0025] In another embodiment, deoxycytidine, deoxythymidine, form B, at least one A fixed-dose powder pharmaceutical composition comprising a residual solvent and at least one pharmaceutically acceptable carrier. The composition is provided, and at least one residual solvent is selected from ICH Class 2 and Class 3 solvents. The concentration of at least one residual solvent is selected from the group consisting of: The fixed dose powder pharmaceutical composition preferably contains undetectable levels of Class 1 With a solvent, in particular 1,2-dichloroethane.

[0026] In yet another embodiment, a disease or disorder characterized by an imbalanced nucleotide pool is Methods of treating a patient with atopic dermatitis are provided, comprising administering a therapeutically effective amount of a composition described herein to a patient with atopic dermatitis. In a specific embodiment, the disorder is TK Thymidine kinase 2 deficiency is characterized by mutations in the thymidine kinase 2 gene.

[0027] In one embodiment, a composition comprising deoxycytidine Form B and at least one residual solvent is provided. and administering to a subject a composition comprising at least one residual solvent selected from the group consisting of ICH Class 2 and Class 3 hydroxybenzoates. The concentration of at least one residual solvent is selected from the group consisting of three solvents, and the concentration of each residual solvent is within the ICH concentration limit of the solvent. The composition may further comprise at least one pharmaceutically acceptable carrier. The pharmaceutical composition may comprise:

[0028] The method includes the steps of administering to the subject a therapeutically effective amount of a second composition comprising deoxythymidine. The second composition may further comprise at least one pharmaceutically acceptable carrier. In such an embodiment, the first and the combined residual solvent content in the second composition is about 100% of the ICH concentration limit for each residual solvent. Less than 10%.

[0029] In another embodiment, one composition administered comprises deoxycytidine Form B and deoxycytidine. oxythymidine and at least one residual solvent, , the concentration of at least one residual solvent selected from ICH Class 2 and Class 3 solvents The concentration of each residual solvent is less than about 10% of the ICH concentration limit. The composition may be a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. The pharmaceutical composition may be a fixed dose powder composition. [Brief explanation of the drawings]

[0030] [Figure 1] FIG. 1 shows the X-ray diffraction pattern of a mixture of Form A and Form B deoxycytidine.

[0031] [Figure 2] FIG. 1 shows the X-ray diffraction pattern of deoxycytidine form B.

[0032] [Figure 3] FIG. 1 shows the X-ray diffraction pattern of deoxycytidine form A. DETAILED DESCRIPTION OF THE INVENTION

[0033] I. Forms of deoxycytidine The present invention relates to crystalline forms of deoxycytidine and compositions comprising crystalline forms of deoxycytidine. The product is provided.

[0034] In one embodiment, deoxycytidine form B is provided. The powder X-ray diffraction (XRPD) pattern of deoxycytidine form B is shown in Figure 2. 13.7°, 17.2°, 18.0°, 19.2° and 22.8° (2θ) (±0.2 In a more specific embodiment, the compound is characterized by an XRPD pattern including a peak at 100° (2θ). and Form B was 13.7°, 17.2°, 18.0°, 19.2° and 22.8° ( It is characterized by an XRPD pattern containing peaks at ±0.1° (2θ). is preferably CuK α1 It is done using radiation.

[0035] The XRPD pattern for Form B is 11.5°, 11.8°, 20.2°, 21.1°, 2 One or more peaks at 1.4° and 21.8° (2θ) (±0.2° (2θ)) In a more specific embodiment, the XRPD pattern may further include 11.5°, 1 1.8°, 20.2°, 21.1°, 21.4° and 21.8° (2θ) (±0.1° (2θ)).

[0036] In specific embodiments, deoxycytidine form B is at 11.5°, 11.8°, 13.7°, 17.2°, 18.0°, 19.2°, 20.2°, 21.1°, 21.4 XRP containing peaks at 21.8°, 21.8° and 22.8° (2θ) (±0.2° (2θ)) In an even more specific embodiment, the deoxyribonucleotides of Form B are characterized by a deoxyribonucleotide pattern D. Jin: 11.5°, 11.8°, 13.7°, 17.2°, 18.0°, 19.2°, 20.2°, 21.1°, 21.4°, 21.8° and 22.8° (2θ) (±0.1 It is characterized by an XRPD pattern containing peaks at 100° (2θ).

[0037] In one embodiment, Form B deoxycytidine is provided in substantially pure form. "Substantially pure," as used herein, means about 95% by weight or more of Form B, about 9 ... 6% by weight or more of Form B, about 97% by weight or more of Form B, about 98% by weight or more of Form B, or about It refers to a mixture of forms of deoxycytidine containing 99% or more by weight of Form B.

[0038] Form B deoxycytidine is also provided in isolated form, i.e., 100% Form B. It can be done.

[0039] Form B deoxycytidine contains minimal residual solvents from manufacture.

[0040] In one embodiment, deoxycytidine Form B is present in less than about 100 ppm of ethyl acetate. , for example, less than about 90 ppm, less than about 80 ppm, less than about 60 ppm, or less than about 50 ppm. In another embodiment, deoxycytidine form B contains 1 ppm or less of ethyl acetate. to about 100 ppm ethyl acetate, for example, 1 ppm to about 90 ppm, 1 ppm to about 80 ppm m, 1 ppm to about 60 ppm, and 1 ppm to about 50 ppm of ethyl acetate.

[0041] Deoxycytidine Form B is preferably dissolved in less than about 100 ppm of n-heptane, preferably less than about 90 ppm of n-heptane. Less than ppm, less than about 80 ppm, less than about 70 ppm, less than about 60 ppm, less than about 50 ppm In another embodiment, the mixture contains less than about 40 ppm, or less than about 30 ppm n-heptane. In this case, deoxycytidine Form B is dissolved in 1 ppm to about 100 ppm of n-heptane, e.g. 1 ppm to approximately 90 ppm, 1 ppm to approximately 80 ppm, 1 ppm to approximately 70 ppm, 1 ppm ~ about 60 ppm, 1 ppm ~ about 50 ppm, 1 ppm ~ about 40 ppm, and 1 ppm ~ about Contains 30 ppm of n-heptane.

[0042] Deoxycytidine Form B is preferably prepared in the presence of less than about 200 ppm ethanol, preferably less than about 150 ppm ethanol. In another embodiment, deoxycytidine form B contains less than 10 ppm of ethanol. 1 ppm to about 200 ppm of ethanol, for example, 1 ppm to about 150 ppm, 1 ppm to Approximately 125ppm, 1ppm to approximately 100ppm, 1ppm to approximately 75ppm, 1ppm to approximately 5 Contains 0 ppm and 1 ppm to about 25 ppm of ethanol.

[0043] In a specific embodiment, deoxycytidine Form B contains less than about 100 ppm acetic acid. ethyl, less than about 100 ppm n-heptane, and less than about 200 ppm ethanol Form B deoxycytidine contains 1 ppm to about 100 ppm ethyl acetate, 1 ppm to about 100 ppm ethyl acetate, May contain about 100 ppm n-heptane and 1 ppm to about 200 ppm ethanol In another embodiment, deoxycytidine Form B is present in an amount of (a) 1 ppm to about 100 ppm m ethyl acetate, (b) 1 ppm to about 100 ppm n-heptane, or (c) 1 pp ethanol from about 200 ppm to about 200 ppm.

[0044] In another specific embodiment, deoxycytidine Form B is present in an amount of less than about 70 ppm acetic acid. ethyl acetate, less than about 50 ppm n-heptane, and less than about 200 ppm ethanol. Form B deoxycytidine contains 1 ppm to about 70 ppm ethyl acetate, 1 ppm to about It may contain 50 ppm n-heptane and 1 ppm to about 200 ppm ethanol. In one embodiment, deoxycytidine Form B is prepared by: (a) administering to a patient in the presence of 1 ppm to about 70 ppm of acetic acid; (b) 1 ppm to about 50 ppm of n-heptane, or (c) 1 ppm to about 2 ppm of n-heptane 00 ppm ethanol.

[0045] Deoxycytidine Form B has a shelf life of at least about 6 months when stored, for example, in a sealed container at room temperature. More specifically, deoxycytidine form B is stable for at least about 12 months. The compound is stable for approximately 18 months or at least 18 months.

[0046] "Stability," as used herein, refers to the stability of XRP Form B of a material after a given storage time. This can be determined by checking the D pattern. , the conversion of form B deoxycytidine to form A deoxycytidine exhibits instability. Stability is assessed by confirming compliance with test attribute criteria as outlined in Example 4. Test attributes include physical appearance, IR trace, active substance and HPLC traces of impurities, water content, residual solvent content, elemental impurities and microbial growth were included. One or more of these attributes may be used over a given period, e.g., 6 months, 12 months, or longer and under various conditions, e.g., 25±2°C / 60±5% relative humidity The temperature can be measured after storage at 40±2°C / 75±5% relative humidity.

[0047] A method for preparing Form B of deoxycytidine is provided in Example 1.

[0048] In one embodiment, the method for preparing Form B deoxycytidine comprises: (a) preparing crude deoxycytidine; (b) contacting the crude deoxycytidine with ethanol. (c) distilling ethanol from the first mixture. (d) contacting the residue with purified water to form a second mixture. (e) heating the second mixture to an internal temperature of at least about 40° C. (f) adding ethanol, ethyl acetate, and heptane to the second mixture. (g) cooling the third mixture to obtain Form B deoxyriboflavin. (h) forming cytidine crystals; and (i) mixing the crystals of form deoxycytidine B with a third mixture. and separating the object.

[0049] The method involves administering more than about 75 kg, e.g., more than about 100 kg, more than about 125 kg, more than about 150 kg, or or for large-scale manufacturing, such as for producing batches of deoxycytidine greater than approximately 200 kg. obtain.

[0050] Crude deoxycytidine is a mixture of polymorphic forms, Form A and Form B, in amounts equal to approximately 100%. In one embodiment, the crude deoxycytidine may contain at least about 5% Form B. , at least about 10% Form B, at least about 20% Form B, at least about 30% Form B Form B, at least about 40% Form B, at least about 50% Form B, at least about 60% Form B, at least about 70% Form B, at least about 80% Form B or at least In another embodiment, the crude deoxycytidine comprises at least about 10% of Form B. 5% Form A, at least about 10% Form A, at least about 20% Form A, at least about 30% Form A, at least about 40% Form A, at least about 50% Form A, at least At least about 60% Form A, at least about 70% Form A, at least about 80% Form A, or contains at least about 10% Form A.

[0051] The distillation in step (c) is preferably carried out at a temperature not exceeding about 70°C, for example about 40°C. The reaction is carried out at a temperature of from about 150°C to less than about 70°C.

[0052] The amount of water used in (d) is preferably about 0.10 to about 0.5 volumes, for example 0 0.25 volume.

[0053] The internal temperature of the second mixture in (e) is preferably about 40°C to about 50°C.

[0054] The solvent in the adding step (f) can be added in any order, either all at once or It can be added in multiple portions. In one embodiment, ethanol is added first. Then, a mixture of ethanol, ethyl acetate, and n-heptane is added. The amounts of ethanol, ethyl acetate, and n-heptane can vary. The amount of ethyl acetate can be 1 volume, 2 volumes, or 3 volumes. The amount of n-heptane can be 1 volume, 2 volumes, or 3 volumes. Solvent or mixture of solvents The material is generally added dropwise or slowly enough to prevent crash precipitation. will be done.

[0055] The third mixture in (g) may be cooled in a stepwise or gradient manner. In this step, the third mixture is cooled to an internal temperature of about 20°C to about 30°C, and the mixture is stirred. In a more specific embodiment, the temperature is maintained at a temperature of The third mixture is stirred for at least about 1 hour, at least about 3 hours, at least about 6 hours, or for at least about 1 hour. The mixture is stirred for at least about 12 hours, or at least about 24 hours, and then further cooled. After stirring, the mixture may be further cooled to an internal temperature of about 0°C to about 10°C.

[0056] The separation step (h) can be carried out by any suitable separation means, such as centrifugal filtration, gravity filtration or vacuum. This can be done by filtration.

[0057] The method provides a composition comprising substantially pure Form B deoxycytidine.

[0058] The crystals of Form B obtained by the method herein contain at least one residual solvent. It may contain and therefore be classified as a composition.

[0059] The composition may contain, in addition to Form B, one or more polymorphic forms of deoxycytidine, e.g., Form It may further contain Form A or other polymorphic forms, including hydrated polymorphic forms. wherein the composition comprises up to about 10% by weight of one or more other polymorphic forms of deoxyribonucleotides. Cytidine, for example, about 0.5% by weight to about 10% by weight, about 1% by weight to about 10% by weight, about 1% by weight % to about 5% by weight or about 1% to about 3% by weight of one or more other polymorphic forms Contains oxycytidine.

[0060] The composition comprising deoxycytidine Form B prepared by the methods described herein comprises: Contains at least one residual solvent. The solvent is classified as an ICH Class 2 (i.e., solvents to be restricted). Class 2 solvents can belong to either Class 1 (i.e., solvents with low toxicity) or Class 2 (i.e., solvents with low toxicity). The maximum permitted daily intake (PDE) and concentration limits are shown in the table below. Class 3 solvents are considered to be less toxic and present a lower risk to human health. The ICH guidelines for Class 3 solvents are 5,000 ppm or 50 mg / day or more. Below.

[0061] TIFF2025160233000001.tif203170

[0062] Class 1 solvents are solvents to be avoided. ICH guidelines state that the solvents are toxic or have unacceptable toxicity. stated that Class 1 solvents should not be used in the manufacture of drug substances due to their harmful environmental impacts. However, if unavoidable, they are permitted in limited amounts. The ICH concentration limits for solvents are shown in the table below.

[0063] TIFF2025160233000002.tif46170

[0064] The methods of manufacture described herein may be obtained, for example, from commercial chemical suppliers or from previously known chemical suppliers. The present invention provides a deoxycytidine composition having a significantly lower residual solvent concentration than a conventional production method. In fact, the method is (a) Class 2 and Class 3 acceptable by ICH guidelines; (b) undetectable levels of ICH Class 3 residual solvents at concentrations / amounts of approximately 10% or less; Deoxycytidine compositions containing a first solvent (particularly 1,2-dichloroethane) are provided. It is optimized to do so.

[0065] "Undetectable," as used herein, means a level that cannot be measured using standard validated methods. Such methods refer to concentrations of a substance that are below the detection limit for that substance when the concentration is determined to be below the detection limit for that substance. are well known in the art and are described, for example, in the United States Pharmacopeia (USP) chapter on residual solvents (467) and and the gas chromatography (GC) method specified in the European Pharmacopoeia (Ph.Eur). The detection limit for 1,2-dichloroethane is, for example, 0.2 ppm.

[0066] In one embodiment, the composition comprises deoxycytidine Form B and at least one residue. Solvents, for example, one residual solvent, two residual solvents, three residual solvents, four residual solvents, five residual solvents or six or more residual solvents.

[0067] In one embodiment, the composition comprises deoxycytidine Form B and at least one residue. and at least one residual solvent is selected from ICH Class 2 and Class 3 solvents. The concentration of at least one residual solvent is selected from the group consisting of: In a more specific embodiment, the composition comprises deoxycytidine form B. and at least one residual solvent, is selected from ICH Class 2 and Class 3 solvents and has a concentration of at least one residual solvent. The concentration is less than about 10% of the ICH concentration limit for the specific residual solvent.

[0068] Preferred Class 2 and Class 3 residual solvents include methanol, toluene, and methylene chloride. ethanol, tert-butyl methyl ether, acetone, ethyl acetate and n-hexane Contains butane.

[0069] Methanol has a permissible daily intake (PDE) of 30 mg or 3,000 ppm. The compositions of the present invention are ICH Class 2 solvents with concentration limits of 1 ppm to about 30 ppm. 0 ppm, for example, 1 ppm to about 250 ppm, 1 ppm to about 200 ppm, 1 ppm to about 150 ppm, 1 ppm to approximately 100 ppm, 1 ppm to approximately 50 ppm, or 1 ppm to approximately It may contain methanol at a concentration of 25 ppm.

[0070] Toluene has a PDE of 8.9 mg / day or a concentration limit of 890 ppm under the ICH The composition of the present invention contains 1 ppm to about 89 ppm, for example, 1 ppm to about 89 ppm. Toluene at concentrations of about 45 ppm, 1 ppm to about 25 ppm, and 1 ppm to about 10 ppm may also contain

[0071] Methylene chloride (dichloromethane) has a PDE of 6.0 mg / day or a concentration of 600 ppm. The compositions of the present invention are ICH Class 2 solvents with a concentration of 1 ppm to about 60 ppm. pm, for example, 1 ppm to about 50 ppm, 1 ppm to about 40 ppm, 1 ppm to about 30 ppm m, 1 ppm to about 20 ppm, 1 ppm to about 10 ppm, and 1 ppm to about 5 ppm It may contain methylene chloride.

[0072] Ethanol has a PDE of 50 mg / day or a concentration limit of 5,000 ppm. The composition of the present invention is a solvent of CH Class 3. pm~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1pp and ethanol at concentrations of 1 ppm to about 100 ppm and 1 ppm to about 50 ppm. .

[0073] tert-Butyl methyl ether (TBME) is a PDE or 5.0 mg / day The compositions of the present invention are ICH Class 3 solvents with a concentration limit of 100 ppm. pm to about 500 ppm, for example, 1 ppm to about 400 ppm, 1 ppm to about 300 ppm, 1 ppm to approximately 200 ppm, 1 ppm to approximately 100 ppm, and 1 ppm to approximately 50 ppm It may also contain a concentration of tert-butyl methyl ether.

[0074] Acetone has a PDE of 50 mg / day or an IC limit of 5,000 ppm. The composition of the present invention is a solvent of H Class 3. m~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1ppm It may contain acetone at a concentration of from about 100 ppm and from 1 ppm to about 50 ppm.

[0075] Ethyl acetate has a PDE of 50 mg / day or a concentration limit of 5,000 ppm. The composition of the present invention is a solvent of CH Class 3. pm~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1pp and ethyl acetate at concentrations of 1 ppm to about 100 ppm and 1 ppm to about 50 ppm. .

[0076] n-Heptane is an ICH Class 1 hazardous substance with a PDE of 50 mg or a limit of 5,000 ppm. The composition of the present invention contains 1 ppm to about 500 ppm, for example, 1 ppm to about 500 ppm. Approx. 400ppm, 1ppm ~ approx. 300ppm, 1ppm ~ approx. 200ppm, 1ppm ~ approx. It may contain n-heptane at a concentration of 100 ppm and 1 ppm to about 50 ppm.

[0077] In a specific embodiment, the composition comprises deoxycytidine form B and a compound of class 2 and / or Class 3 ICH solvents (a) 1 ppm to about 300 ppm methanol, ( b) toluene from 1 ppm to about 89 ppm, (c) methylene chloride from 1 ppm to about 60 ppm (d) 1 ppm to about 500 ppm of ethanol; (e) 1 ppm to about 500 ppm of T BME, (f) acetone from 1 ppm to approximately 500 ppm, (g) acetone from 1 ppm to approximately 500 ppm ethyl acetate, and (h) 1 ppm to about 500 ppm n-heptane Also includes one.

[0078] 1,2-Dichloroethane (1,2-DCE) is an ICH hazardous substance with a concentration limit of 5 ppm. The composition of the present invention is a Class 1 solvent. pm, preferably less than 0.2 ppm, of 1,2-DCE, and therefore 1,2-DCE is undetectable.

[0079] A pharmaceutical composition comprising deoxycytidine Form B provided herein and at least one residual solvent. The composition may have a shelf life of at least 6 months, e.g., at least 9 months, at least 12 months, or The stability of the composition was determined according to Example 4, i.e., for a period of at least 24 months. , can be measured according to confirming compliance with test attributes when subjected to storage conditions .

[0080] In another embodiment, there is provided deoxycytidine form A. The XRPD pattern of deoxycytidine is shown in Figure 3. Form A of deoxycytidine has an XRPD of 8.4°, 12. 9°, 14.2°, 16.8°, 18.4°, 19.4°, 28.4°, 30.0° and The XRPD pattern was characterized by peaks at 30.6° (2θ) and 30.6° (±0.2° (2θ)). In more specific embodiments, Form A has an angle of 8.4°, 12.9°, 14.2°, 16.8°, 18.4°, 19.4°, 28.4°, 30.0° and 30.6° (2θ ) (±0.1° (2θ)). XRPD is Preferably CuK α1 It is done using radiation.

[0081] The XRPD pattern of Form A shows peaks at 21.3°, 21.6° and 22.2° 2θ (± 0.2° (2θ)). In the XRPD pattern, the peaks are at 21.3°, 21.6° and 22.2° (2θ) (± 0.1° (2θ)).

[0082] In one embodiment, Form A deoxycytidine is provided in substantially pure form. "Substantially pure," as used herein, means about 95% by weight or more of Form A, about 9% by weight or more of Form B, about 9% by weight or more of Form C, about 9% by weight or more of Form D, about 9% by weight or more of Form A, about 9% by weight or more of Form B ... 6% by weight or more of Form A, about 97% by weight or more of Form A, about 98% by weight or more of Form A or about Refers to a mixture of forms of deoxycytidine containing 99% or more by weight of Form A.

[0083] II. Pharmaceutical Compositions The present invention also provides a method for treating a disease comprising administering to a subject a subject, comprising administering to a subject a method for treating a disease, the method ... The present invention provides a pharmaceutical composition comprising the compound of formula (I) and at least one pharmaceutically acceptable carrier.

[0084] In one embodiment, the pharmaceutical composition comprises substantially pure Form B deoxyriboflavin as the active ingredient. cycytidine, at least one residual solvent, and at least one pharmaceutically acceptable carrier. and,

[0085] In another embodiment, the pharmaceutical composition comprises substantially pure Form B deoxycytidine and and deoxythymidine, where deoxythymidine also acts as an active ingredient. the mixture, at least one residual solvent, and at least one pharmaceutically acceptable carrier; Includes:

[0086] In yet another embodiment, the pharmaceutical composition comprises deoxycysteine ​​in Form B as described herein. The present invention also includes a composition comprising a gin and at least one pharmaceutically acceptable carrier.

[0087] In yet another embodiment, the pharmaceutical composition comprises deoxycysteine ​​in Form B as described herein. a composition comprising deoxythymidine, a composition comprising deoxythymidine, and at least one pharmaceutically acceptable salt thereof and a carrier on which the

[0088] Compositions comprising deoxythymidine contain at least 97% by weight of deoxythymidine, e.g. For example, about 97-99% by weight deoxythymidine or 98-99% by weight deoxythymidine The remainder was one or more impurities, including at least one residual solvent. That's fine.

[0089] Such pharmaceutical compositions comprise a therapeutically effective amount of deoxycytidine Form B (and, where applicable, and a therapeutically effective amount of deoxythymidine if desired), at least one pharmaceutically acceptable carrier; The phrase "pharmaceutically acceptable" means physiologically tolerable and typically When administered to humans, it may cause allergic or similar adverse reactions, such as stomach upset or dizziness. non-toxic, approved by federal or state regulatory agencies, or Specifically, for human use, the United States Pharmacopoeia or other generally recognized pharmacy "Carrier" refers to a molecular entity or composition that is administered with a therapeutic agent. A pharmaceutical carrier is a sterile liquid. Bodies, e.g., physiological saline solutions in water or oil, e.g., of petroleum, animal, vegetable, or synthetic origin For example, peanut oil, soybean oil, mineral oil, sesame oil, etc. When the pharmaceutical composition is administered intravenously, When used in conjunction with a sachet, saline solution is the preferred carrier. Aqueous glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions.

[0090] Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, Malt, rice, wheat flour, chalk, silica gel (anhydrous silica colloid), magnesium stearate Sodium, Sodium Stearate, Glycerol Monostearate, Talc, Sodium Chloride Contains ethanol, dried skim milk, glycerol, propylene, glycol, water, etc. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Good too.

[0091] The pharmaceutical composition may include at least one glidant. The glidant may be added during the manufacturing process. A flow agent is a substance that counteracts the poor flowability of the powder in the container. Colloidal silicon dioxide, e.g., AEROSIL, magnesium trisilicate (M g), powdered cellulose, starch, talc and tricalcium phosphate, Not limited to:

[0092] The pharmaceutical composition may include at least one lubricant. A lubricant reduces interparticle friction. Exemplary lubricants include magnesium stearate, aluminum stearate, Aluminum (Al) or calcium stearate, PEG 4000-8000 and talc, hard Hydroxylated castor oil, stearic acid and its salts, glycerol esters and stearyl fumarate These include, but are not limited to, sodium carbonate, and hydrogenated cottonseed oil.

[0093] In a specific embodiment, the fixed dose pharmaceutical composition comprises deoxycytidine Form B and deoxycytidine. cythymidine, at least one residual solvent, at least one fluidizing agent, and at least and one lubricant, and the composition is in the form of a powder.

[0094] The amount of deoxycytidine Form B in the pharmaceutical composition is from 1,000 mg to about 5,000 mg. , for example, about 1,000 mg to about 4,000 mg, about 1,000 mg to about 3,000 mg Alternatively, it may be about 1,000 mg to about 2,000 mg.

[0095] The amount of deoxythymidine in the pharmaceutical composition is 1,000 mg to about 5,000 mg, for example , about 1,000 mg to about 4,000 mg, about 1,000 mg to about 3,000 mg, or about It may be 1,000 mg to about 2,000 mg.

[0096] In a preferred embodiment, both deoxycytidine and deoxythymidine in form B is present at approximately 2,000 mg.

[0097] The ratio of deoxycytidine to deoxythymidine can vary. For example, they can be 50 / 5 0 ratio, or approximately 5 / 95, 10 / 90, 15 / 85, 20 / 80, 25 / 75, 30 / 70, 35 / 65, 40 / 60, 45 / 55, 55 / 45, 60 / 40, 65 / 35, 7 Ratios of 0 / 30, 75 / 25, 80 / 20, 85 / 15, 90 / 10, and 95 / 5 It is also possible.

[0098] At least one fluidizing agent is present in an amount of from about 0.1% to about 10% by weight, for example, from about 1% to about 5% Alternatively, the fluidizing agent may be present in an amount of about 1% to about 3%. Silicon dioxide (e.g., AEROSIL 200).

[0099] At least one lubricant is present in an amount of from about 0.1% to about 1% by weight, for example from about 0.1% to about 0. It may be present in an amount of 5% or about 0.3% to about 0.7%. The agent is magnesium stearate.

[0100] Deoxycytidine Form B is from about 40% to about 60% by weight, for example from about 40% to about 50% by weight, about 50% by weight to about 60% by weight, or about 45% by weight to about 55% by weight. There can be.

[0101] Similarly, deoxythymidine is about 40% by weight to about 60% by weight, for example, about 40% by weight to about 50% by weight, about 50% by weight to about 60% by weight, or about 45% by weight to about 55% by weight. There can be.

[0102] The powdered pharmaceutical composition may be packaged in any suitable form, for example, a bottle, a pouch or a sachet. In a preferred embodiment, the pharmaceutical composition is made of laminated PET, aluminum and The powdered pharmaceutical composition is then packaged in a pouch made of low-density polyethylene. An oral solution is prepared by dissolving in water.

[0103] At least one residual solvent is deoxycytidine Form B (as described above), a pharmaceutically acceptable salt thereof Carry-over from the manufacture of either the acceptable carrier or deoxythymidine, if applicable. obtain.

[0104] The pharmaceutical composition may contain at least one residual solvent, for example, two residual solvents, three residual solvents, Contains four or more residual solvents.

[0105] At least one residual solvent is selected from ICH Class 2 and Class 3 solvents, The concentration of at least one residual solvent is less than approximately 10% of the ICH concentration limit for each residual solvent. .

[0106] Preferred Class 2 and Class 3 residual solvents include methanol, toluene, and methylene chloride. ethanol, tert-butyl methyl ether, acetone, ethyl acetate and n-hexane Contains butane.

[0107] Methanol has a permissible daily intake (PDE) of 30 mg or 3,000 ppm. The pharmaceutical composition of the present invention is an ICH Class 2 solvent with a concentration limit of 1 ppm to about 300 ppm, e.g. 1 ppm to about 250 ppm, 1 ppm to about 200 ppm, 1 ppm ~150 ppm, 1 ppm ~ 100 ppm, 1 ppm ~ 50 ppm or 1 ppm It may contain methanol at a concentration of up to about 25 ppm.

[0108] Toluene has a PDE of 8.9 mg / day or a concentration limit of 890 ppm under the ICH The pharmaceutical composition of the present invention contains 1 ppm to about 89 ppm, for example 1 ppm to about 45 ppm, 1 ppm to about 25 ppm, and 1 ppm to about 10 ppm. It may contain ene.

[0109] Methylene chloride (dichloromethane) has a PDE of 6.0 mg / day or a concentration of 600 ppm. The pharmaceutical composition of the present invention is an ICH Class 2 solvent with a concentration of 1 ppm to about 6 ppm. 0 ppm, for example, 1 ppm to about 50 ppm, 1 ppm to about 40 ppm, 1 ppm to about 30 ppm, 1 ppm to approximately 20 ppm, 1 ppm to approximately 10 ppm, and 1 ppm to approximately 5 ppm The solution may contain methylene chloride at a concentration of

[0110] Ethanol has a PDE of 50 mg / day or a concentration limit of 5,000 ppm. The pharmaceutical composition of the present invention contains 1 ppm to about 500 ppm, for example, 1ppm~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1 Even if it contains ethanol at concentrations of 1 ppm to about 100 ppm and 1 ppm to about 50 ppm good.

[0111] tert-Butyl methyl ether (TBME) is a PDE or 5.0 mg / day The pharmaceutical composition of the present invention is an ICH Class 3 solvent with a concentration limit of 100 ppm. 1 ppm to about 500 ppm, for example, 1 ppm to about 400 ppm, 1 ppm to about 300 ppm m, 1 ppm to approximately 200 ppm, 1 ppm to approximately 100 ppm, and 1 ppm to approximately 50 ppm It may contain tert-butyl methyl ether at a concentration of m.

[0112] Acetone has a PDE of 50 mg / day or an IC limit of 5,000 ppm. The pharmaceutical composition of the present invention contains 1 ppm to about 500 ppm, for example, 1 ppm~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1p may contain acetone at concentrations of from 1 ppm to about 100 ppm and from 1 ppm to about 50 ppm. .

[0113] Ethyl acetate has a PDE of 50 mg / day or a concentration limit of 5,000 ppm. The pharmaceutical composition of the present invention contains 1 ppm to about 500 ppm, for example, 1ppm~about 400ppm, 1ppm~about 300ppm, 1ppm~about 200ppm, 1 ppm to about 100 ppm and 1 ppm to about 50 ppm of ethyl acetate. good.

[0114] n-Heptane is an ICH Class 1 hazardous substance with a PDE of 50 mg or a limit of 5,000 ppm. The pharmaceutical composition of the present invention contains 1 ppm to about 500 ppm, for example, 1 ppm ~400ppm, 1ppm~300ppm, 1ppm~200ppm, 1ppm~ It may contain n-heptane at a concentration of about 100 ppm and 1 ppm to about 50 ppm.

[0115] In specific embodiments, the pharmaceutical compositions described herein comprise: (a) 1 ppm to about 30 0 ppm methanol, (b) 1 ppm to approximately 89 ppm toluene, (c) 1 ppm to approximately 60 ppm methylene chloride, (d) 1 ppm to approximately 500 ppm ethanol, (e) 1 p pm to approximately 500 ppm TBME, (f) 1 ppm to approximately 500 ppm acetone, (g) 1 ppm to about 500 ppm of ethyl acetate, and (h) 1 ppm to about 500 ppm of n- Heptane.

[0116] In another specific embodiment, the pharmaceutical composition comprises deoxycytidine Form B and deoxycytidine. cythymidine and at least one pharmaceutically acceptable carrier; and (a) 1 ppm to about 300 ppm of cythymidine. ppm of methanol, (b) 1 ppm to about 89 ppm of toluene, (c) 1 ppm to about 6 0 ppm methylene chloride, (d) 1 ppm to approximately 500 ppm ethanol, (e) 1 ppm m to approximately 500 ppm TBME, (f) 1 ppm to approximately 500 ppm acetone, (g) 1 ppm to about 500 ppm of ethyl acetate, and (h) 1 ppm to about 500 ppm of n-hexane and at least one of:

[0117] 1,2-Dichloroethane (1,2-DCE) is an ICH hazardous substance with a concentration limit of 5 ppm. It is a Class 1 solvent. The composition of the present invention contains 0.2 ppm to about 0.5 ppm, preferably It may contain 1,2-DCE at a concentration of less than 0.2 ppm, and therefore 1,2-DCE is undetectable.

[0118] Oral administration is the preferred method of administration. The active ingredient is extracted from both bovine and human udders. Any food consumed by the patient, including but not limited to milk, infant formula, and water The composition may be added to a liquid in the form of a granule.

[0119] Additionally, pharmaceutical compositions suitable for oral administration may be in the form of capsules, tablets, powders, granules, solutions, syrups, or the like. It may be in the form of a capsule, a suspension (in a non-aqueous or aqueous liquid), or an emulsion. Hard gelatin capsules are made with lactose, starch or its derivatives, magnesium stearate, Sodium, saccharin sodium, cellulose, magnesium carbonate, stearic acid or The soft gelatin capsule may contain vegetable oils, waxes, fats, semi-solid or liquid polymers. Solutions and syrups may contain water, polyols and sugars. The active agent intended for administration is a drug that delays disintegration and / or absorption of the active agent in the gastrointestinal tract. The drug may be coated with or mixed with a material that allows for sustained release. can be achieved over an extended period of time, and if necessary, the active agent can be protected from degradation in the stomach. Pharmaceutical compositions for oral administration may be suitable for use in certain gastrointestinal locations due to specific pH or enzymatic conditions. The active agent may be formulated to enhance release of the active agent at the site.

[0120] Intrathecal administration is the more preferred form of administration to overcome the problem of crossing the blood / brain barrier. Intrathecal administration is the process of delivering a drug into the spinal canal, more specifically into the spinal cord, so that it reaches the cerebrospinal fluid. This involves injection into the subthecal space. This method is commonly used for spinal anesthesia, chemotherapy, and analgesics. Intrathecal administration is by lumbar puncture (bolus injection) or port-catheter system. This can be done by stem (bolus or infusion). Catheters are the most common It is inserted between the lumbar lamina and the tip penetrates the medullary cavity to the desired level (generally L3-L4). Intrathecal formulations most commonly use water and saline as vehicles, but EDTA and lipids have also been used.

[0121] A more preferred mode of administration is parenteral administration, including intravenous administration. Pharmaceutical compositions adapted for oral administration may contain antioxidants, buffers, bacteriostats, and other additives that may be useful in treating the composition. Aqueous and non-aqueous sterile injection solutions or suspensions which may contain solutes that make them substantially isotonic with blood Other ingredients that may be present in such compositions include water, alcohol, polio Compositions adapted for parenteral administration may be packaged in sealed envelopes. They may be provided in single-dose or multi-dose containers such as bottles and vials, which must be sterilized immediately prior to use. It can be stored in a freeze-dried (lyophilized) form, requiring only the addition of a bacterial carrier. Injectable solutions and suspensions can be prepared from sterile powders, granules, and tablets. Suitable vehicles that can be used to provide the form are well known to those skilled in the art. Water for Injection USP; Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose Injection Aqueous vehicles such as sucrose and sodium chloride injection, lactated Ringer's injection, etc.; ethyl alcohol Water-miscible vehicles such as chol, polyethylene glycol and polypropylene glycol Corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate and non-aqueous vehicles such as propyl and benzyl benzoate.

[0122] Additionally, some patients may have received enteral nutrition before deoxynucleoside treatment was initiated. Because of the potential for dN to be administered via a gastric feeding tube or other enteral feeding means, It is possible.

[0123] Further modes of administration include mucosal administration, such as nasal, sublingual, vaginal, buccal, or rectal administration; This includes transdermal administration to a subject.

[0124] Pharmaceutical compositions adapted for nasal and pulmonary administration may include solid carriers such as powders. It may be administered by rapid inhalation through the nose. Compositions for nasal administration may be sprays or Alternatively, inhalation directly into the lungs can be achieved by inhaling deeply. This can be achieved by introduction through a syringe or through a mouthpiece. The composition may comprise an aqueous or oily solution of the active ingredient. Compositions for inhalation may be administered by pressurized aerosol. Specially adapted devices, including but not limited to sols, nebulizers or inhalers The active ingredient may be supplied in a dosage form which may be configured to provide a predetermined dose of the active ingredient.

[0125] Pharmaceutical compositions adapted for rectal administration may be presented as suppositories or enemas. Suitable pharmaceutical compositions include pessaries, tampons, creams, gels, pastes, and foams. Alternatively, it may be provided as a spray formulation.

[0126] Pharmaceutical compositions adapted for transdermal administration are intended to be in intimate contact with the epidermis of the recipient for a prolonged period of time. It may be provided as a separate patch intended to maintain

[0127] III. Treatment Methods The present invention also provides a therapeutically effective amount of a composition described herein comprising deoxycytidine Form B. The method of claim 1, further comprising administering to the subject a method for treating an imbalanced nucleotide in a subject in need thereof. Methods for treating diseases or disorders characterized by pools are provided.

[0128] In some embodiments, the method further comprises treatment with a second composition comprising deoxythymidine. The method further comprises the step of administering an effective amount to a subject.

[0129] In another embodiment, one nucleotide sequence containing both deoxycytidine and deoxythymidine is The composition is administered.

[0130] In one embodiment, a composition comprising deoxycytidine Form B and at least one residual solvent is provided. and administering to a subject a composition comprising at least one residual solvent selected from the group consisting of ICH Class 2 and Class 3 hydroxybenzoates. The concentration of at least one residual solvent is selected from the group consisting of three solvents, and the concentration of each residual solvent is within the ICH concentration limit of the solvent. It is less than about 10% of the value.

[0131] In a more specific embodiment, deoxycytidine form B and at least one A composition consisting of residual solvents is administered to a subject, and at least one residual solvent is administered. The solvent is selected from ICH Class 2 and Class 3 solvents and contains at least one residual solvent. The concentration is less than about 10% of the ICH concentration limit of each residual solvent. Any composition may be administered.

[0132] In a specific embodiment, the composition comprises deoxycytidine Form B and (a) 1 ppm (b) toluene from 1 ppm to about 89 ppm; (c) toluene from 1 ppm to about 300 ppm; pm to about 60 ppm methylene chloride, (d) 1 ppm to about 500 ppm ethanol, ( e) 1 ppm to approximately 500 ppm of TBME, (f) 1 ppm to approximately 500 ppm of acetone , (g) 1 ppm to about 500 ppm of ethyl acetate, and (h) 1 ppm to about 500 ppm of ethyl acetate. and at least one of m n-heptanes.

[0133] In a preferred embodiment, the ICH Class 1 solvent, particularly 1,2-dichloroethane, is Undetectable in the composition.

[0134] In another embodiment, deoxycytidine Form B, at least one residual solvent, and and a pharmaceutically acceptable carrier as described herein, and the residual solvent is , the concentration of at least one residual solvent selected from ICH Class 2 and Class 3 solvents is less than approximately 10% of the ICH concentration limit for each residual solvent.

[0135] In a more specific embodiment, deoxycytidine, deoxythymidine, thiamin, and thiamin forms B and C are at least one residual solvent and at least one pharmaceutically acceptable solvent as described herein A pharmaceutical composition consisting of a carrier is administered, and residual solvents are classified as ICH Class 2 and and Class 3 solvents, and the concentration of at least one residual solvent is selected from the I In a specific embodiment, the pharmaceutical composition has a CH concentration of less than about 10% of the CH concentration limit. is a fixed dose powder composition.

[0136] In yet another embodiment, a disease or disorder characterized by an imbalanced nucleotide pool is A method for treating a disease or disorder in a subject in need of such treatment comprises: (1) administering to a subject a compound of formula (I) of Form B; (2) a first composition comprising oxycytidine and at least one residual solvent; a therapeutically effective second composition comprising oxythymidine and optionally at least one residual solvent; and administering to a subject an amount of at least one residual solvent that meets ICH Class 2 and and Class 3 solvents, and the concentration of at least one residual solvent is selected from the IC In such an embodiment, the sum of (1) and (2) is less than about 10% of the H concentration limit. The combined residual solvent amount is less than about 10% of the ICH concentration limit for that particular residual solvent.

[0137] In a specific embodiment, the first composition (i.e., (1)) and / or the second composition The composition (i.e., (2)) is a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier. It is a composition.

[0138] Deoxycytidine and deoxythymidine are administered in the same or separate pharmaceutical compositions The ratio of deoxycytidine to deoxythymidine can vary, for example, whether or not , they are in a 50 / 50 ratio, or about 5 / 95, 10 / 90, 15 / 85, 20 / 80 , 25 / 75, 30 / 70, 35 / 65, 40 / 60, 45 / 55, 55 / 45, 60 / 40, 65 / 35, 70 / 30, 75 / 25, 80 / 20, 85 / 15, 90 / 10, and and a 95 / 5 ratio.

[0139] Diseases characterized by imbalanced nucleotide pools that can be treated by the methods of the present invention The disorder involves the following genes: TK2; DGUOK; TYMP; RRM2B; SUCLA2 and those characterized by mutations in MPV17. In a preferred embodiment, the disorder includes, but is not limited to, mitochondrial DNA damage. In a more preferred embodiment, the MDS is a TK2 depletion syndrome. Myopathy characterized by mutations in SUCLA2, and encephalopathies characterized by mutations in SUCLA3. Myopathy type, neurogastrointestinal encephalopathy type characterized by mutations in TYMP, and DGUOK, This includes hepatopathic disorders characterized by mutations in POLG, and MPV17. In a preferred embodiment, the disorder is thymidine kinase, characterized by a mutation in the TK2 gene. This is enzyme 2 deficiency.

[0140] Administration of the pharmaceutical composition is directed to treating disorders characterized by imbalanced nucleotide pools, such as It should be initiated as soon as MDS is suspected and continued throughout the patient's life. Tests for the diagnosis of such disorders, including deficiencies, are known in the art.

[0141] For example, dT and dC are administered in equal amounts in a mixture for TK2 deficiency. Selection of an effective dose will be determined by one of ordinary skill in the art, taking into consideration several factors known to those skilled in the art. Such factors include specific forms of deoxynucleosides, as well as bioavailability. These include its pharmacokinetic parameters such as irritability, metabolism, and half-life, which are established during the normal development procedures typically used in obtaining regulatory approval for a drug compound. Additional factors to consider when considering dosage include the condition or disease being treated or the normal The benefit achieved in the body, the weight of the patient, the route of administration (whether administration is acute or chronic), (regardless of the cause), concomitant medications, and medications known to affect the efficacy of the administered drug. Certain other factors are involved, therefore, the precise dose will depend on the judgment of the artisan and each patient's circumstances. The clinical outcome should be determined according to standard clinical techniques.

[0142] A preferred dose ranges from about 100 mg / kg / day to about 1,000 mg / kg / day. A more preferred dose is in the range of about 200 mg / kg / day to about 800 mg / kg / day. A more preferred dose is in the range of about 250 mg / kg / day to about 400 mg / kg / day. These dosages may be used for individual deoxynucleosides or for two or more deoxynucleosides. The dosage of a composition containing a mixture of thiazolinone, e.g., dT and dC. For example, the dose is In another example, the dose may include 200 mg / kg, 400 mg / kg / day of dT alone. In another example, the dose may include a mixture of 200 mg / kg / day dT and 200 mg / kg / day dC. , 400 mg / kg / day of a mixture of dT and dC.

[0143] Deoxynucleosides are administered once daily, twice daily, three times daily, four times daily, or five times daily. , up to six times a day, preferably on a regular basis. If administered four times daily, dosing should occur at 8:00 AM, 12:00 PM, 4:00 PM, and and it will be 8:00 PM.

[0144] If administered intravenously or intrathecally, the dose may be lower.

[0145] A preferred dose range for such administration is from about 50 mg / kg / day to about 500 mg / kg / day. kg / day.

[0146] Dosage can be adjusted to optimize efficacy in a subject. The synucleoside was initiated at 100 mg / kg / day and then administered as directed to assess the subject's response and tolerability. Depending on tolerance, 200 mg / kg / day, 400 mg / kg / day, 800 mg / kg / day, The dose may be increased over time up to a maximum of 1000 mg / kg / day.

[0147] Before increasing the dosage, the subject may be monitored for improvement of their condition. The subject's response to therapeutic administration of the creoside is evaluated by measuring the subject's muscle strength and control, mobility, etc. These parameters can be monitored by observing changes in body mass and height. If one or more of these parameters increase after administration, treatment can be continued. If one or more of the meters stays the same or decreases, the deoxynucleoside The dosage can be increased.

[0148] Deoxynucleosides may be co-administered with other drugs. Such drugs include M These include therapeutic agents for treating the symptoms of certain forms of DS, particularly TK2 deficiency. , dT and dC are tetrahydrouridine (an inhibitor of cytidine deaminase) and iodopropanol. Ammocillin H (an inhibitor of purine nucleoside phosphorylase) and tipiracil (thymidylidene A wide range of enzyme inhibitors, including but not limited to, inhibitors of phosphodiesterase and phosphodiesterase These drugs may be co-administered with inhibitors of endogenous nucleoside catabolic enzymes. It is known to be effective in treating some cancers. [Example]

[0149] Example 1: Preparation of deoxycytidine form B

[0150] Deoxycytidine Form B was produced by the following 130 kg engineering process.

[0151] Step 1: The concentrated residue (crude deoxycytidine) from reactor R-6424 is removed by vacuum. By passing it through the filter housing, 924 kg of water is discharged through a dedicated pipe. of ethanol (99.5%) was added.

[0152] Step 2: The reactants in reactor R-6426 were stirred for approximately 30 minutes.

[0153] Step 3: The reactant in reactor R-6426 was distilled under reduced pressure at 70°C or less.

[0154] Step 4: The concentrated residue in reactor R-6426 was vented using nitrogen. By passing through the housing, 98L (0.25 volume) of purified water is passed through a dedicated pipe. did.

[0155] Step 5: The reactants in reactor R-6426 were stirred for approximately 10 minutes.

[0156] Step 6: The internal temperature of reactor R-6426 was increased to about 48°C.

[0157] Step 7: Maintain internal temperature and stir until mixture is visibly dissolved.

[0158] Step 8: Reactor R-6425 was placed under vacuum. 308 kg (0.70 vol) of ethanol was added. The ethanol (99.5%) was introduced through a dedicated pipe and then released using a nitrogen vacuum. Ta.

[0159] Step 9: The internal temperature of reactor R-6426 was maintained between 40°C and 50°C. Pressurized ethanol (99.5%) in a container R-6425 was heated under nitrogen to prevent crystal precipitation. The mixture was slowly added to reactor R-6426 using a 1000 vol.

[0160] Step 10: 616 kg (1.58 vol) of ethanol (99.5%), 702 kg (1.8 vol) ethyl acetate and 730 kg (1.36 vol) heptane were reacted under vacuum. Charged into reactor R-6425. Vacuum was released using nitrogen. Mixture was stirred.

[0161] Step 11: The internal temperature of the reactor R-6426 was maintained between 40°C and 50°C. The pressurized mixture of -6425 was slowly added to reactor R-6426.

[0162] Step 12: The internal temperature of reactor R-6426 was cooled to between 20°C and 30°C.

[0163] Step 13: The mixture was stirred for 1 hour.

[0164] Step 13: The internal temperature of reactor R-6426 was cooled to between 0°C and 10°C.

[0165] Step 14: The mixture was stirred for about 1 hour.

[0166] Step 15: The internal temperature of reactor R-6426 was maintained between 0°C and 10°C.

[0167] Step 16: The crystals were collected using a centrifugal filter and washed with ethyl acetate. By D analysis, the crystals were characterized as deoxycytidine Form B (see Example 2). (I want to be).

[0168] Example 2: Lot test results

[0169] The production of dC on a 4 kg scale resulted in Form B of dC (Figure 2). Pull-up resulted in a mixture of Form A and Form B dC (Figure 1), with the remainder of the final API. The distillate content increased.

[0170] Lot 2DC(7) / SP-17001 was manufactured at a 4 kg scale.

[0171] Lots 2DC(7)-6-18001, 2DC(7)-6-18002 and and 2DC(7)-6-18003 were 55 kg production lots. , which were prepared by a method different from that described in Example 1 herein, and which are shown in Figure 1 as Form A and Form B. A mixture of A and B was produced.

[0172] Lots 2DC(7)-E-6-19001 and 2DC(7)-E-6-19002 are Using the process described in Example 1, 130 kg of the product was prepared to provide the Form B polymorph (Figure 2). It was a manufacturing operation.

[0173] TIFF2025160233000003.tif244170

[0174] Mixture of A and B dC isoforms (2DC (A+B mix)), small-scale production The B isoform (2DC(7) / SP-17001 early stage) generated by the operation, and XRPD data for the A isoform isolated in Table 2 are shown. By determining the XRPD after (2DC(7) / SP-17001 12 months) , and stability form B was confirmed.

[0175] TIFF2025160233000004.tif124170

[0176] Example 3: Residual Solvent Content of Commercially Available Materials

[0177] Samples of dC and dT purchased from a commercial supplier (Carbosynth) were Residual solvent content was determined by MS analysis, and the results are shown in the table below:

[0178] TIFF2025160233000005.tif60170

[0179] Example 4: Stability Study of Deoxycytidine Form B

[0180] Stability of Form B deoxycytidine prepared by the method of Example 1 over a 12-month period Tests were conducted to determine the effectiveness of the material over time. Four lots of material were tested: (1) and ( 2) is packed in a polyethylene (PE) bag sealed with a cable tie, and (3) is packed in a cable (4) is packed in a PE bag containing silica gel and sealed with a lid. Packaged in E drums.

[0181] The following test attributes were measured: Appearance (white to off-white powder) IR tracing HPLC ○Purity 99.0% or more Less than 1.0% total impurities ○0.5% or less cytosine Less than 0.05% α-anomer ●Water content 0.5% or less ●Specific rotation (+57.0°~+60.0°) ●Ignition residue ○0.5% or less ●Residual solvent (GC) ○MeOH: 3,000ppm or less Toluene: 890 ppm or less Methylene chloride: 600 ppm or less ○MTBE: 5,000ppm or less Ethyl acetate: 5,000 ppm or less n-heptane 5,000 ppm or less Ethanol: 5,000 ppm or less Elemental impurities ○Cd: 5ppm or less ○Pb: 5ppm or less ○As: 15ppm or less ○Hg:30ppm or less ○Co:50ppm or less ○V: 100ppm or less ○Ni: 200ppm or less ○Ti: 8ppm or less ○Au: 100ppm or less ○Pd: 100ppm or less ○Ir: 100ppm or less ○Os: 100ppm or less ○Rh: 100ppm or less ○Ru: 100ppm or less ○Se: 150ppm or less ○Ag: 150ppm or less ○Pt: 100ppm or less ○Mo: 3,000ppm or less ○Cu: 3,000ppm or less ○Cr: 11,000ppm or less ●Microbial content ○ Total aerobic microbial count: 100 CFU / g or less Total yeast and mold count: 100 CFU / g or less

[0182] Two sets of conditions were applied: (1) long-term storage (25±2°C / 60±5% relative humidity) and and (2) accelerated storage (40±2°C / 75±5% relative humidity).

[0183] All attributes were measured initially (0 months) to ensure compliance with the above criteria. Compliance was confirmed at 6, 9 and 12 months by visual, IR, HPLC and moisture content measurements. Microbial content was measured again at 12 months. No significant changes were observed from baseline to 12 months.

[0184] Example 5: Fixed-dose pharmaceutical composition

[0185] Contains 2'-deoxycytidine (2.0 g) and 2'-deoxythymidine (2.0 g) A fixed dose powder for oral solution was prepared in the amounts shown in the table below. The powder is laminated with polyethylene terephthalate, aluminum and low density polyethylene. The oral solution was packaged in a foil pouch (stick pack) consisting of 100mg of PEG-1000. The powder was prepared by dissolving the ingredients in water.

[0186] TIFF2025160233000006.tif74170

Claims

1. A composition comprising deoxycytidine of form B and at least one residual solvent, Deoxycytidine of form B is characterized by a powder X-ray diffraction (XRPD) pattern containing peaks at 13.7°, 17.2°, 18.0°, 19.2°, and 22.8°(2θ) (±0.2°(2θ)). Deoxycytidine in form B is, The first mixture is produced by contacting crude deoxycytidine with ethanol, Distilling ethanol from the first mixture to produce a residue, The aforementioned residue is brought into contact with purified water to produce a second mixture, The internal temperature of the second mixture is heated to at least about 40°C, Adding ethanol, ethyl acetate, and heptane to the second mixture to produce a third mixture, The third mixture is cooled to produce deoxycytidine of form B, Obtained by a method including, The at least one residual solvent is selected from the group consisting of methanol, toluene, methylene chloride, ethanol, tert-butyl methyl ether, acetone, ethyl acetate, and n-heptane, and the concentration of the at least one residual solvent is (a) methanol at a concentration of 1 ppm to about 300 ppm, (b) toluene at a concentration of 1 ppm to about 89 ppm, (c) methylene chloride at a concentration of 1 ppm to about 60 ppm, (d) ethanol at a concentration of 1 ppm to about 500 ppm, (e) tert-butyl methyl ether (TBME) at a concentration of 1 ppm to about 500 ppm, (f) acetone at a concentration of 1 ppm to about 500 ppm, (g) ethyl acetate at a concentration of 1 ppm to about 500 ppm, and (h) n-heptane at a concentration of 1 ppm to about 500 ppm. The composition is stable for at least three months at 25±2°C and 60±5% relative humidity. composition.

2. The deoxycytidine of the form B further comprises one or more XRPD peaks at 11.5°, 11.8°, 20.2°, 21.1°, 21.4°, and 21.8°2θ±0.2°2θ, The composition according to claim 1.

3. A solution comprising less than 100 ppm of ethyl acetate, less than 100 ppm of n-heptane, and less than 200 ppm of ethanol. The composition according to claim 1.

4. A solution comprising less than 70 ppm of ethyl acetate, less than 50 ppm of n-heptane, and less than 200 ppm of ethanol. The composition according to claim 1.

5. The composition comprises at least one of the following: (a) methanol at a concentration of 1 ppm to about 300 ppm, (b) toluene at a concentration of 1 ppm to about 89 ppm, (c) methylene chloride at a concentration of 1 ppm to about 60 ppm, (d) ethanol at a concentration of 1 ppm to about 500 ppm, (e) tert-butyl methyl ether (TBME) at a concentration of 1 ppm to about 500 ppm, (f) acetone at a concentration of 1 ppm to about 500 ppm, (g) ethyl acetate at a concentration of 1 ppm to about 500 ppm, and (h) n-heptane at a concentration of 1 ppm to about 500 ppm. The composition according to claim 1.

6. 1,2-dichloroethane is undetectable, The composition according to claim 1.

7. Further comprising at least one pharmaceutically acceptable carrier, The composition according to any one of claims 1 to 6.

8. Further comprising deoxythymidine, The composition according to any one of claims 1 to 6.

9. A fixed-dose pharmaceutical composition comprising deoxycytidine of form B, deoxythymidine, at least one residual solvent, and at least one pharmaceutically acceptable carrier, a. The at least one residual solvent is selected from the group consisting of methanol, toluene, methylene chloride, ethanol, tert-butyl methyl ether, acetone, ethyl acetate, and n-heptane, and the concentration of the at least one residual solvent is less than 10% of the ICH concentration limit for each residual solvent; b. The composition is in powder form; c. The weight ratio of deoxycytidine to deoxythymidine is 50:

50. The composition is stable for at least three months at 25±2°C and 60±5% relative humidity. Fixed-dose pharmaceutical composition.

10. The at least one pharmaceutically acceptable carrier comprises at least one fluidizing agent and at least one lubricant, The composition according to claim 9.

11. The at least one fluidizing agent is present in an amount of about 0.1% by weight to about 10% by weight. The composition according to claim 10.

12. The at least one fluidizing agent comprises colloidal silicon dioxide, The composition according to claim 11.

13. The at least one lubricant is present in an amount of about 0.1% by weight to about 1% by weight. The composition according to claim 10.

14. The at least one lubricant comprises magnesium stearate, The composition according to claim 13.

15. Deoxycytidine is present in an amount of about 40% to about 50% by weight. The composition according to claim 10.

16. The powder is packaged in a pouch, The composition according to claim 10.

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