Metal-polyphenol complex particles, drug-lipid particles, and methods for producing the same and their applications

Abstract

The present disclosure relates to the field of biopharmaceutical technology, specifically to metal-polyphenol complex particles, drug-lipid particles, and their manufacturing methods and applications. The present disclosure provides a metal-polyphenol complex as a drug carrier for drug stability, delivery, and transport, etc., and forms metal-polyphenol complex particles with other carriers, which act in concert to achieve effective administration of negatively charged drugs. While achieving efficient systemic drug delivery, toxicity is significantly reduced compared to LNPs containing cationic or ionizable lipids, enabling safe and effective treatment of diseases or conditions.

Claims

1. The use of metal-polyphenol complexes in the manufacture of nucleic acid delivery systems, The aforementioned metal-polyphenol complex is formed by the reaction of a polyphenol molecular portion and a metal ion portion, and the polyphenol molecular portion and the metal ion portion are linked by a coordination bond. The metal ions of the metal-polyphenol complex are bound to the nucleic acids via coordination bonds. The aforementioned polyphenol molecular portion is one or more selected from curcumin, hesperetin, catechin, and epigallocatechin gallate. The aforementioned metal ion portion is Fe ３＋ Ca ２＋ Mg ２＋ Al ３＋ A combination of one or more types, selected from the others, for use.

2. The aforementioned polyphenol molecular portion is selected from curcumin, hesperetin, catechin, and epigallocatechin gallate. The aforementioned metal ion portion is Fe ３＋ Ca ２＋ , and Al ３＋ A more selected use according to claim 1.

3. The polyphenol molecular portion is selected from curcumin (formula 1), hesperetin (formula 5), ​​catechin (formula 13), and epigallocatechin gallate (formula 18). 【Chemistry 1】 【change】 The use described in claim 2.

4. The use according to any one of claims 1 to 3, wherein the molar ratio of the polyphenol molecular portion to the metal ion portion is 1:(0.5 to 2).

5. The polyphenol molecular portion is curcumin (formula 1), and the metal ion portion is Fe ３＋ And, Curcumin (Formula 1) and Fe ３＋ have a molar ratio of 1:1, or The polyphenol molecular portion is curcumin (formula 1), and the metal ion portion is Al ３＋ And, Curcumin (Equation 1) and Al ３＋ The use according to claim 4, wherein the molar ratio is 1:

1.

6. The nucleic acids delivered by the nucleic acid delivery system are one or more types selected from mRNA, siRNA, sgRNA, ASO, circRNA, microRNA, DNA, ecDNA, and artificial nucleic acids. The nucleic acid delivery system is used to introduce nucleic acids into cells. The nucleic acid is used to silence the expression of a target sequence in a mammalian subject, or to treat a mammalian disease or condition. The aforementioned mammal is a human, The disease or condition is related to gene expression, and the gene includes a drug target sequence. The aforementioned diseases or conditions include cancer, viral infections, autoimmune diseases, diabetes, or Alzheimer's disease. The aforementioned viral infections include hepatitis A, hepatitis B, hepatitis C, SARS-CoV-2, HIV, HPV, influenza, smallpox, or syphilis. The aforementioned cancers include liver cancer, glioma, melanoma, lung cancer, pancreatic cancer, or breast cancer. The nucleic acid delivery system is used in the manufacture of vaccines. The use according to any one of claims 1 to 5, wherein the vaccine is a COVID-19 vaccine.

7. Metal-polyphenol composite particles, The aforementioned metal-polyphenol composite particles are (i) A metal-polyphenol complex comprising a polyphenol molecular portion and a metal ion portion, linked by a coordination bond, The aforementioned polyphenol molecular portion is one or more selected from curcumin, hesperetin, catechin, and epigallocatechin gallate. The aforementioned metal ion portion is Fe ３＋ Ca ２＋ Mg ２＋ , and Al ３＋ A combination of one or more types that are selected from among them. (ii) A conjugation lipid that inhibits particle aggregation, wherein the conjugation lipid that inhibits particle aggregation contains a PEG-lipid conjugate and / or PEG-DAA, (iii) Metal-polyphenol complex particles containing non-cationic lipids or non-ionizable lipids other than conjugation lipids that inhibit particle aggregation.

8. The aforementioned polyphenol molecular portion is selected from curcumin, hesperetin, catechin, and epigallocatechin gallate. The aforementioned metal ion portion is selected from Fe³⁺, Ca²⁺, and Al³⁺. The PEG-lipid conjugate is phosphatidylethanolamine-polyethylene glycol 2000 (Formula 47), phosphatidylethanolamine-polyethylene glycol 700 (Formula 48), phosphatidylethanolamine-polyethylene glycol 1000 (Formula 49), phosphatidylethanolamine-polyethylene glycol 5000 (Formula 50), and one or more combinations selected from these. R1 and R2 are both independently as follows: 【Chemistry 2】 【change】 【change】 【change】 【change】 The non-cationic lipid or non-ionizable lipid described in (iii) is lecithin PC, phosphatidylethanolamine PE, phosphatidylserine PS, phosphatidic acid PA, phosphatidylglycerol PG, ceramide-1-phosphate CP, phosphatidylinositol PI, phosphatidylthreonine PT, sphingomyelin SM, lysolecithin LPC, lysophosphatidylethanolamine LPE, lysophosphatidylserine LPS, lysophosphatidic acid LPA, lysophosphatidylglycerol LPG, lysophosphatidylinositol LPI, lysophosphatidylthreonine LPT, lysosphingomyelin LSM, sphingosine-1-phosphate S1P, and one or more combinations selected therefrom, as described in claim 7.

9. The metal-polyphenol complex particle according to claim 8, wherein the polyphenol molecular portion is selected from curcumin (formula 1), hesperetin (formula 5), ​​catechin (formula 13), and epigallocatechin gallate (formula 18).

10. The metal-polyphenol composite particle according to claim 8, wherein the PEG-lipid conjugate is one or more types selected from DSPE-PEG2000, DSPE-PGE700, DSPE-PEG1000, and DSPE-PEG5000.

11. The metal-polyphenol composite particle according to claim 10, wherein the PEG-lipid conjugate is one or more selected from DSPE-PEG2000 (formula 58), DSPE-PGE700 (formula 55), DSPE-PEG1000 (formula 56), and DSPE-PEG5000 (formula 57). 【Transformation 3】 【change】

12. (iii) The non-cationic lipids or non-ionizable lipids described are lecithin (PC) (Formula 29), phosphatidylethanolamine (PE) (Formula 30), phosphatidylserine (PS) (Formula 31), phosphatidic acid (PA) (Formula 32), phosphatidylglycerol (PG) (Formula 33), ceramide-1-phosphate (CP) (Formula 34), phosphatidylinositol (PI) (Formula 35), phosphatidylthreonine (PT) (Formula 36), sphingomyelin (SM) (Formula 37), lysolecithin (LPC) (Formula 38), lysophosphatidylethanolamine (LPE) (Formula 39), lysophosphatidylserine (LPS) (Formula 40 The metal-polyphenol complex particle according to claim 8, wherein R1 and R2 are one or more selected from lysophosphatidic acid (LPA) (formula 41), lysophosphatidylglycerol (LPG) (formula 42), lysophosphatidylinositol (LPI) (formula 43), lysophosphatidylthreonine (LPT) (formula 44), lysosphingomyelin (LSM) (formula 45), and sphingosine-1-phosphate (S1P) (formula 46), and R1 and R2 are both independently a decanoyl group, a lauroyl group, a palmitoyl group, a stearoyl group, an oleoyl group, a linoleoyl group, an ercoyl group, an arachidoyl group, or a diphytanoyl group. 【Chemistry 4】 【change】 【change】 【change】 【change】 【change】

13. The metal polyphenol complex particle according to any one of claims 8 to 12, wherein the non-cationic lipid or non-ionizable lipid described in (iii) further comprises cholesterol.

14. The metal polyphenol complex particles according to claim 13, wherein the non-cationic lipid or non-ionizable lipid described in (iii) comprises cholesterol and one or more combinations selected from DSPC, DSPE, DSPA, and DSPG.

15. The metal polyphenol complex particle according to claim 14, wherein the non-cationic lipid or non-ionizable lipid described in (iii) comprises cholesterol (formula 59) and one or more combinations selected from DSPC (formula 51), DSPE (formula 52), DSPA (formula 53), or DSPG (formula 54). 【Transformation 5】 【change】

16. The metal polyphenol complex particle according to claim 15, wherein the non-cationic lipid or non-ionizable lipid described in (iii) comprises cholesterol (formula 59) and DSPC (formula 51).

17. The metal polyphenol composite particle according to any one of claims 7 to 16, wherein the molar ratio of the polyphenol molecular portion to the metal ion portion is 1:(0.5 to 2).

18. The polyphenol molecular portion is curcumin (formula 1), and the metal ion portion is Fe ３＋ And, Curcumin (Equation 1) and Fe ３＋ The molar ratio of is 1:1, or The polyphenol molecular portion is curcumin (formula 1), and the metal ion portion is Al ３＋ And, Curcumin (Equation 1) and Al ３＋ The metal-polyphenol composite particles according to claim 17, wherein the molar ratio is 1:

1.

19. The metal-polyphenol complex particles according to claim 13, wherein the metal-polyphenol complex particles consist of (i) a metal-polyphenol complex, (ii) a conjugation lipid that inhibits particle aggregation, and (iii) a non-cationic lipid or a non-ionizable lipid, the mole fraction of the metal-polyphenol complex in the raw material is 10% to 20%, the mole fraction of the conjugation lipid that inhibits particle aggregation in the raw material is 2% to 10%, the mole fraction of the cholesterol in the raw material is 0% to 48%, and the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 40% to 75%.

20. The metal-polyphenol complex particles consist of (i) a metal-polyphenol complex, (ii) a conjugation lipid that inhibits particle aggregation, and (iii) a non-cationic lipid or a non-ionizable lipid, wherein the mole fraction of the metal-polyphenol complex in the raw material is 5% or more and less than 10%, the mole fraction of the conjugation lipid that inhibits particle aggregation in the raw material is 2% to 10%, the mole fraction of the cholesterol in the raw material is 0% to 48%, and the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 30% or more and less than 40%, or 40% to 75%, or The metal-polyphenol complex particles according to claim 13, wherein the metal-polyphenol complex particles consist of (i) a metal-polyphenol complex, (ii) a conjugation lipid that inhibits particle aggregation, and (iii) a non-cationic lipid or a non-ionizable lipid, the mole fraction of the metal-polyphenol complex in the raw material is 10% to 20%, the mole fraction of the conjugation lipid that inhibits particle aggregation in the raw material is 2% to 10%, the mole fraction of the cholesterol in the raw material is 0% to 48%, and the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 30% or more and less than 40%.

21. The mole fraction of the metal-polyphenol complex in the raw material is 5% or more and less than 10%, 10% to 15%, or 15% to 20%. The mole fraction of the metal-polyphenol complex in the raw material is 5%, 10%, or 15%. The mole fraction of lipids in the conjugation that inhibits particle aggregation in the raw material is 3% to 5% or 5% to 10%. The mole fraction of lipids in the conjugation that inhibits particle aggregation in the raw material is 3%, 5%, or 10%. The mole fraction of cholesterol in the raw material is 10% to 30%, 30% to 47%, or 10% to 20%. The mole fraction of cholesterol in the raw material is 10%, 30%, or 47%. The mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw materials is 45% to 55%, 60% to 65%, or 50% to 65%. The metal-polyphenol complex particles according to claim 19 or claim 20, wherein the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 45%, 55%, 60%, or 65%.

22. The mole fraction of the metal-polyphenol complex in the raw material is 5% or more and less than 10% or 10% to 15%, the mole fraction of the conjugation lipids that inhibit particle aggregation in the raw material is 5% to 10%, the mole fraction of the cholesterol in the raw material is 10% to 30%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 60% to 65%, and the metal ion portion in the metal-polyphenol complex is Fe ３＋ More selected, The mole fraction of the metal-polyphenol complex in the raw material is 15%, the mole fraction of the conjugation lipids that inhibit particle aggregation in the raw material is 10%, the mole fraction of the cholesterol in the raw material is 10%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 65%, and the metal ion portion in the metal-polyphenol complex is Fe ３＋ More selected, The mole fraction of the metal-polyphenol complex in the raw material is 5%, the mole fraction of the conjugation lipid that inhibits particle aggregation in the raw material is 5%, the mole fraction of the cholesterol in the raw material is 30%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 60%, and the metal ion portion in the metal-polyphenol complex is Fe ３＋ A metal-polyphenol composite particle according to claim 19 or claim 20, more preferably selected.

23. The mole fraction of the metal-polyphenol complex in the raw material is 5% or more but less than 10% or 10%. The mole fraction of the conjugation lipids that inhibit particle aggregation in the raw material is 3% to 5%, the mole fraction of the cholesterol in the raw material is 30% to 47%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 45% to 55%, and the metal ion portion in the metal-polyphenol complex is Al ３＋ More selected, The mole fraction of the metal-polyphenol complex in the raw material is 5%, the mole fraction of the conjugation lipids that inhibit particle aggregation in the raw material is 3%, the mole fraction of the cholesterol in the raw material is 47%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 45%, and the metal ion portion in the metal-polyphenol complex is Al ３＋ To be selected, or The mole fraction of the metal-polyphenol complex in the raw material is 10%, the mole fraction of the conjugation lipids that inhibit particle aggregation in the raw material is 5%, the mole fraction of the cholesterol in the raw material is 30%, the mole fraction of non-cationic lipids or non-ionizable lipids other than cholesterol in the raw material is 55%, and the metal ion portion in the metal-polyphenol complex is Al ３＋ A metal-polyphenol composite particle according to claim 19 or claim 20, more preferably selected.

24. A method for producing metal-polyphenol composite particles according to any one of claims 7 to 23, The metal-polyphenol complex particles are obtained by mixing (i) a metal-polyphenol complex, (ii) a conjugation lipid that inhibits particle aggregation, and (iii) a non-cationic lipid or a non-ionizable lipid, and the method for producing them.

25. Step 1 involves reacting the polyphenol molecule with the metal ion via a coordination bond to form a metal-polyphenol complex. Step 2 involves mixing the metal-polyphenol complex produced in Step 1 with a conjugation lipid that inhibits particle aggregation, a non-cationic lipid, or a non-ionizable lipid to produce the metal-polyphenol complex particles. Includes, Polyphenol molecules are dissolved in ethanol, and then metal ions are added and reacted to obtain the metal-polyphenol complex. The molar ratio of polyphenol molecules to metal ions is 1:(1-2), The manufacturing method according to claim 24, wherein the conditions for the reaction include reacting at 60°C for 1 hour.

26. Drug-lipid particles, wherein the drug-lipid particles are (a) A drug, which is a negatively charged molecule, (b) comprising metal-polyphenol composite particles according to any one of claims 7 to 23, The drug is encapsulated within the metal-polyphenol complex particles. The drug is one or more types selected from nucleic acids, proteins, polypeptides, small molecules, nucleic acid analogs, protein analogs, and polypeptide analogs, The nucleic acid is a drug-lipid particle, which is one or more types selected from mRNA, siRNA, sgRNA, ASO, circRNA, microRNA, DNA, ecDNA, or artificial nucleic acids.

27. A method for producing drug-lipid particles according to claim 26, wherein the drug is encapsulated in the metal-polyphenol complex particles to obtain the drug-lipid particles.

28. (a) a drug, (i) a metal-polyphenol complex, (ii) a conjugation lipid that inhibits particle aggregation, and (iii) a non-cationic lipid or a non-ionizable lipid, Obtain the drug-lipid particles, A metal-polyphenol complex, a conjugation lipid that inhibits particle aggregation, and a non-cationic or non-ionizable lipid are dissolved in an organic compound to form an organic phase, a drug is dissolved in a buffer to form an aqueous phase, and the organic phase and aqueous phase are uniformly mixed to obtain drug-lipid particles. The aforementioned organic compound is ethanol. The aforementioned buffer is an enzyme-free Tris-HCl buffer. The manufacturing method according to claim 27, wherein the mixing method of the organic phase and the aqueous phase includes a microfluidic chip or ultrasound.

29. Use of metal-polyphenol complex particles according to any one of claims 7 to 23 or drug-lipid particles according to claim 26 in the manufacture of a composition used for drug delivery.

30. The aforementioned composition is used to introduce a drug into cells. The composition is a drug. The aforementioned drugs are used to silence the expression of target sequences in mammalian subjects, to deliver drugs within the mammalian body, to deliver drugs from the body to mammalian cells, or to treat mammalian diseases or conditions. The aforementioned mammal is a human, The disease or condition is related to gene expression, and the gene includes a drug target sequence. The aforementioned diseases or conditions include cancer, viral infections, autoimmune diseases, diabetes, or Alzheimer's disease. The aforementioned viral infections include hepatitis A, hepatitis B, hepatitis C, SARS-CoV-2, HIV, HPV, influenza, smallpox, or syphilis. The aforementioned cancers include liver cancer, glioma, melanoma, lung cancer, pancreatic cancer, or breast cancer. The aforementioned drug is a vaccine, The use according to claim 29, wherein the route of administration of the drug includes intrathecal injection, intramuscular injection, intracranial injection, intravenous injection, or intratumor injection.

31. A drug containing metal-polyphenol complex particles according to any one of claims 7 to 23 or drug-lipid particles according to claim 26.

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