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CN-122003446-A - GMP (good manufacturing practice) large-scale production method for synthesizing glucan-guanidine-bisphosphonate conjugate by one-pot method

CN122003446ACN 122003446 ACN122003446 ACN 122003446ACN-122003446-A

Abstract

The present invention relates to a large scale GMP synthesis process for modified dextran conjugates, more specifically to a dextran-guanidine-bisphosphonate conjugate, wherein the bisphosphonate is alendronate. The large scale GMP synthesis method of the present invention enables the production of high purity, pharmaceutical grade dextran-guanidine-bisphosphonate conjugates. The invention also relates to the use of the dextran-guanidine-bisphosphonate conjugate in a medicament for the treatment of tumors.

Inventors

  • A. R. Holmberg

Assignees

  • 德克斯特克医疗公司

Dates

Publication Date
20260508
Application Date
20241010
Priority Date
20231013

Claims (14)

  1. 1. A method of synthesizing a glucan-guanidine-bisphosphonate conjugate, the method comprising the sequential steps of: (a) Adding periodate to an aqueous solution containing dextran followed by sulfuric acid; (b) Adding sodium dihydrogen phosphate and then sodium hydroxide into the solution obtained in the step (a); (c) Adding alendronate to the solution obtained in step (b); (d) Adding aminoguanidine to the solution obtained in step (c); (e) Adding a sodium borohydride solution to the solution obtained in step (d) to produce a dextran-guanidine-bisphosphonate conjugate; (f) Purifying the glucan-guanidine-bisphosphonate conjugate obtained in step (e), preferably by tangential flow filtration, to obtain a purified glucan-guanidine-bisphosphonate conjugate; (g) Diluting the purified dextran-guanidine-bisphosphonate conjugate obtained in step (f) with sodium acetate and sodium chloride to produce a diluted dextran-guanidine-bisphosphonate conjugate, adjusting the pH of the diluted dextran-guanidine-bisphosphonate conjugate to 6.0-7.0, and filtering the diluted dextran-guanidine-bisphosphonate conjugate.
  2. 2. The method according to claim 1, characterized in that the bisphosphonate is an aminobisphosphonate, preferably an alendronate-based.
  3. 3. The method according to claim 1 or 2, wherein in step (a) the pH is adjusted to a pH of 1.5-pH 1.9, preferably to a pH of 1.55-pH 1.8, more preferably to a pH of 1.6-pH 1.7.
  4. 4. A method according to any of the preceding claims, wherein in step (b) the pH is adjusted to pH 7-pH 7.7, preferably pH 7.3-pH 7.7, more preferably pH 7.3-pH 7.5.
  5. 5. The method according to any of the preceding claims, wherein step (a), step (c) or both step (a) and step (c) are stirred under light-protected conditions.
  6. 6. The method according to any of the preceding claims, wherein step (d), step (e) or both step (d) and step (e) are stirred under light-protected conditions.
  7. 7. The method according to any one of the preceding claims, wherein the dextran-guanidine-bisphosphonate conjugate in step (g) is filtered through a bacterial entrapment filter.
  8. 8. A method according to any of the preceding claims, characterized in that the method is a large scale one-pot GMP process.
  9. 9. A dextran-guanidine-bisphosphonate conjugate obtained by the method of any of the preceding claims, characterized in that one molecule of the dextran-guanidine-bisphosphonate conjugate comprises 5 to 8 alendronate molecules and 40 to 45 aminoguanidine molecules.
  10. 10. A glucan-guanidine-bisphosphonate conjugate, characterized in that one molecule of the glucan-guanidine-bisphosphonate conjugate comprises 5 to 8 alendronate molecules and 40 to 45 aminoguanidine molecules.
  11. 11. A pharmaceutical formulation, characterized in that it comprises a dextran-guanidine-bisphosphonate conjugate according to claim 8 or 9, and a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable adjuvant.
  12. 12. The glucan-guanidine-bisphosphonate conjugate of claim 8 or 9 or the pharmaceutical formulation of claim 10 for use in the treatment of cancer.
  13. 13. The glucan-guanidine-bisphosphonate conjugate or the pharmaceutical formulation of claim 11, wherein the cancer is a cancer located in bone.
  14. 14. The glucan-guanidine-bisphosphonate conjugate or the pharmaceutical formulation according to claim 11 or 12, characterized in that the cancer is selected from cancer metastasis to bone or cancer with bone as primary site, preferably the cancer is selected from multiple myeloma, metastatic castration resistant prostate cancer (mCRPC), bone metastatic lung cancer and bone metastatic breast cancer.

Description

GMP (good manufacturing practice) large-scale production method for synthesizing glucan-guanidine-bisphosphonate conjugate by one-pot method Technical Field The present invention relates to a method for synthesizing modified glucan conjugates, and more particularly, to a large-scale GMP synthesis method that can obtain pharmaceutical grade glucan-guanidine-bisphosphonate (bissphonate) conjugates. The invention also relates to the use of the dextran-guanidine-bisphosphonate conjugate in a medicament for the treatment of tumors. Background Dextran-guanidine-bisphosphonate conjugates are a candidate drug for the treatment of cancer metastasis at bones with increased bone cell activity. The conjugates showed good safety and therapeutic efficacy in clinical studies (phase I and II) without the occurrence of serious adverse events associated with the drug (Thellenberg-Karlsson et al 2023). Holmberg et al 2010 and Holmberg et al 2009 disclose the biological effects of dextran-guanidine-bisphosphonate conjugates. The present inventors' patents EP2131867B1 and EP2274018B1 describe the function and properties of dextran-guanidine-bisphosphonate conjugates, as well as the micro-preparation of said conjugates. Clinical studies of dextran-guanidine-bisphosphonate conjugates require a large scale manufacturing process that meets good manufacturing practice (Good Manufacturing Practice, GMP). GMP methods refer to process flows designed, developed, and performed in accordance with GMP standards. GMP is a set of regulations, guidelines and procedures mandated by regulatory authorities (e.g., FDA, EMA in europe) to ensure that products, particularly pharmaceuticals, medical devices and foods, are consistently produced and controlled and meet quality standards. The distinction between laboratory synthesis and Good Manufacturing Practice (GMP) synthesis includes scale, regulatory requirements, equipment control and documented grades. The large-scale production of drug candidates is challenging and complex. This is a multi-step complex process involving technical and chemical challenges, while also requiring an increase in the dosing amounts of reactants (API, pharmaceutically active ingredients). It requires careful consideration, balancing and optimization of API concentration, reaction kinetics, safety, equipment design, API quality, cost, regulatory compliance, and the like. Problems that occur in large-scale production typically do not occur in small scale and may not be predictable. The GMP production process of successful drug candidates has little resemblance to the original laboratory synthesis process of the invention, leaving only the principle and product in agreement. Development of such GMP production processes may take years. The transfer from laboratory scale to GMP scale typically fails, e.g. good experimental results cannot be reproduced in a transfer from a small scale development stage to GMP standard scale. Due to chemical/technical limitations and restrictions that cannot be exceeded, research drugs may not be mass produced under GMP standards at all. The transition from preclinical to clinical research in drug development is subject to stringent regulatory requirements, particularly in the production of drug candidates. It requires a large-scale GMP synthesis process that can obtain pharmaceutical grade products. Every subtle element of the synthesis process must be strictly in compliance with GMP specifications. A complete file named test drug declaration Material (IMPD) is provided to the regulatory agency for the regulatory agency to obtain detailed information and approval of the drug. IMPD includes complete chemical and pharmaceutical information, covering drug quality standards, analytical methods, pre-clinical information, and regulatory and legal information. The auditing of IMPD is a critical/mandatory requirement before regulatory approval for developing a clinical trial is obtained. This ensures that the test drug meets GMP standards before entering a clinical trial in a human subject. Disclosure of Invention The object of the present invention is to provide a large scale GMP synthesis process, also an optimized and improved preparation of dextran-guanidine-bisphosphonate conjugates, wherein the bisphosphonate group is preferably alendronate (alendronate group). The present invention achieves the above object by a synthesis process as defined in the independent claims. Preferred embodiments of the invention are described in the dependent claims. The process is a sequential one-pot process comprising the steps of dextran activation, alendronate (alendronate) conjugation, aminoguanidine conjugation and reductive amination followed by purification by tangential flow filtration (TANGENTIAL FLOW FILTRATION, TFF) optimizing the filtration cycle. The prior art method is on the order of mu L/mL and cannot be applied to large-scale (L) synthesis. The present invention can produce high purity pharmaceuti