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CN-122005812-A - Combined medicine composition for engineering modification of in vivo target cells

CN122005812ACN 122005812 ACN122005812 ACN 122005812ACN-122005812-A

Abstract

The invention provides a combination pharmaceutical composition for engineering target cells in vivo, wherein the target cells do not comprise myeloid cells, and the combination pharmaceutical composition comprises a first medicine with active ingredients which can saturate, inhibit or clear substances with phagocytic capacity of the myeloid cells in vivo and a second medicine with active ingredients which can engineer the target cells in vivo. The invention reduces or eliminates endocytosis of the marrow cell line to the target drug before the target cell engineering transformation by combining the marrow cell inhibition drug and the cell engineering transformation drug of the target cell, not only reduces off-target effect, but also improves lymphoid organ targeting, reduces toxic and side effects of the cell engineering transformation drug to liver, and has important significance for in vivo (in situ) lymphocyte engineering transformation (including gene editing) or immunotherapy (such as in vivo CAR T cell therapy).

Inventors

  • CUI YANFANG

Assignees

  • 瑞达信使(杭州)生物技术有限公司

Dates

Publication Date
20260512
Application Date
20241111

Claims (10)

  1. 1. A combination pharmaceutical composition for engineering a target cell in vivo or in situ, wherein the target cell does not comprise a myeloid cell, the combination pharmaceutical composition comprising a first drug and a second drug, the active ingredient of the first drug being a substance capable of saturating, inhibiting or eliminating phagocytic capacity of the myeloid cell in vivo, the active ingredient of the second drug being a nucleic acid molecule, polypeptide, protein, small molecule or epigenetic modulator in combination with a macromolecule capable of engineering a target cell in vivo.
  2. 2. The combination of claim 1, wherein the first and second agents comprise any one of a pharmaceutically acceptable dosage form, the first and second agents being two separate formulations or a single formulation mixed together, the first and second agents being administered sequentially or in parallel.
  3. 3. The combination of claim 1, wherein the substance capable of scavenging or inhibiting phagocytic capacity of myeloid cells in vivo comprises bisphosphonates, diamidines, chloroquine and its derivatives, gadolinium chloride and its derivatives; the diamidine medicine comprises propamidine and derivatives thereof; the bisphosphonates comprise one or more of hydroxyethylphosphonate, clodronate, pamidronate, tiludronate, alendronate, risedronate, ibandronate and zoledronic acid; the substance capable of saturating phagocytic capacity of in vivo marrow cells comprises one or more of allogeneic red blood cell antibody, liposome or other temporary occupation Fc receptor, scavenger receptor, complement receptor, high molecular polymer, poly arginine, polyinosinic acid, silicon-based nanoparticle, heparin and heparinase.
  4. 4. The combination pharmaceutical composition according to claim 1, wherein the nucleic acid molecule comprises DNA and/or RNA, preferably one or more of mRNA, siRNA, ASO, linear DNA or circular plasmid DNA, the polypeptide protein comprises an editing enzyme, and the small molecule compound epigenetic modulator comprises a stem cell differentiation factor.
  5. 5. The combination pharmaceutical composition according to claim 1, wherein the first drug further comprises a first delivery vehicle capable of delivering the active ingredient of the first drug into the body, the first vehicle comprising a viral vehicle, a viroid vehicle or a non-viral vehicle comprising a liposome, micelle, dendrimer, microsphere or microcapsule, the first vehicle preferably being a liposome and a viral vehicle and having a preference for myeloid cells in the body.
  6. 6. The combination pharmaceutical composition according to claim 1, wherein the second drug further comprises a second delivery vehicle capable of targeted delivery of the active ingredient of the second drug to lymphocytes in vivo, the second vehicle comprising a viral-like vehicle, a viroid-like vehicle or a non-viral-like vehicle, the non-viral-like vehicle comprising a liposome, micelle, dendrimer, microsphere or microcapsule, the second vehicle preferably being a liposome and a viral-like vehicle, and the liposome comprising at least one targeting modified lipid compound, the targeting modification comprising an antibody modification capable of binding specifically to an antigen on the surface of a target cell in vivo.
  7. 7. The combination pharmaceutical composition according to claim 1, wherein the first drug and the second drug are pharmaceutically acceptable any one dosage form respectively, the first drug and the second drug further comprise additives, the additives comprise stabilizers and/or diluents, the additives are added in an amount which is 1% -30% of the total mass of the injection, the stabilizers comprise sucrose or trehalose, and the diluents comprise one or more of phosphate buffer solution, acetate buffer solution, citrate and tris hydrochloride buffer solution.
  8. 8. The combination of claim 1, wherein the first and second agents, when injectable, are administered by local intramuscular, subcutaneous, endothelial, intratumoral injection or infusion, respectively, or by intravenous injection.
  9. 9. The combination pharmaceutical composition of claim 1, wherein the in vivo myeloid cells comprise one or more of macrophages and their precursors, mast cells, dendritic cells, neutrophils, basophils, eosinophils; And/or the in vivo target cells comprise in vivo target cells comprising one or more of neurons, stem cells, epithelial cells, endothelial cells, fibroblasts, lymphocytes comprising one or more of T cells, B cells, NK cells, NKT cells.
  10. 10. Use of a combination composition according to any one of claims 1 to 9 for the preparation of an in vivo target cell, such as a lymphocyte engineering preparation or an immunotherapeutic preparation.

Description

Combined medicine composition for engineering modification of in vivo target cells Technical Field The invention belongs to the technical field of in-vivo or in-situ cell engineering transformation, and particularly relates to a combined medicine composition for in-vivo target cell engineering transformation. Background In vivo (in situ) lymphocyte engineering or gene editing, as represented by in vivo CAR T cell therapy, is expected to reduce the cost and toxic side effects of existing conventional CAR T cell therapies. However, because of the natural phagocytic nature of myeloid-derived cells, which are predominantly monocytes, macrophages, dendritic cells, a significant portion of the cell-engineered material, such as DNA, RNA, or proteins, is phagocytosed by these myeloid-derived cells when delivered into the body via a non-viral, viroid vector or viral delivery system in order to generate CAR T in vivo or in situ. Not only is there a potential dose toxicity due to material wastage, but the consequences of decellularized targets are also unexpected, which is one of the reasons that limits CAR T cell therapy development and popularization in vivo. This similar situation also exists for in vivo or in situ engineering of other target cells besides lymphocytes. Disclosure of Invention The invention aims to provide a novel combination composition for engineering in-vivo target cells, which can reduce the probability of off-target of in-situ or in-vivo engineering target cells and improve in-situ or in-vivo target cell targeting and in-situ lymphoid organ targeting. In order to achieve the above object, the present invention provides the following technical solutions: The first aspect of the invention provides a combination pharmaceutical composition for engineering target cells in vivo, wherein the target cells comprise lymphocytes, the combination pharmaceutical composition comprises a first medicament and a second medicament, the active ingredient of the first medicament is a substance capable of saturating, inhibiting or eliminating phagocytic capacity of myeloid cells in vivo, and the active ingredient of the second medicament is an epigenetic regulator capable of engineering nucleic acid molecules, polypeptides, proteins, small molecules or a combination thereof with macromolecules of the target cells in vivo. The target cells include lymphocytes and other target cells other than myeloid lineage cells. In an embodiment of the invention, the first drug and the second drug comprise any one of the pharmaceutically acceptable dosage forms. In the embodiment of the present invention, the first drug and the second drug are two separate formulations or a single formulation after mixing, and the first drug and the second drug are sequentially or concurrently administered. Further, if the second drug is administered after the first drug is administered for a period of time, the length of the interval is not particularly limited, and may be determined based on monitoring of myeloid cells and lymphocytes or other target cells in the body after administration. Further, it is within the scope of the present invention to administer the first and second agents simultaneously or first and then the first agent. Further, it is within the scope of the present invention that the first and second medicaments are mixed formulations. In theory, any substance in the art that is capable of inhibiting, scavenging or saturating phagocytic capacity of myeloid cells in vivo without affecting or with less impact on lymphocytes (or other target cells) can be used as the active ingredient of the first agent. Including new substances with such functions as have been proven and are to be developed by the public. Preferably, the substance that can clear or inhibit phagocytic capacity of myeloid cells in vivo includes bisphosphonates, diamidine drugs, chloroquine and its derivatives, gadolinium chloride and its derivatives. Specifically, the diamidine drugs include propamidine and derivatives thereof. In particular, the bisphosphonates include, but are not limited to, one or more of hydroxyethylphosphonate, clodronate, pamidronate, tiludronate, alendronate, risedronate, ibandronate, zoledronic acid, and other forms of formulation thereof such as liposome-entrapped carriers. The substance capable of saturating phagocytic capacity of marrow cells in vivo comprises allogeneic red blood cell antibody, liposome or other substances which can temporarily occupy Fc receptor, scavenger receptor or complement receptor, etc., and have low toxicity such as high molecular polymer, poly arginine, polyinosinic acid or silicon-based nanoparticle, etc., and also comprises heparin or heparinase, etc. Preferably, the nucleic acid molecule comprises DNA and/or RNA, preferably one or more of mRNA, siRNA, ASO, linear DNA or circular plasmid DNA. Preferably, the polypeptide protein comprises an editing enzyme, such as cas9, cas12, and the like. Preferably, the