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CN-121971639-A - Medicine for treating urinary system diseases

CN121971639ACN 121971639 ACN121971639 ACN 121971639ACN-121971639-A

Abstract

A medicine for treating urinary system diseases is provided. Specifically provided is a complex comprising a carrier protein selected from the group consisting of a mutant DIIIbV of the subfraction structure of the albumin third domain (e.g., DIIIb), an albumin third domain DIIIV containing the mutant, and a multimer of said DIIIbV or DIIIV, and a functional molecule for use in the diagnosis, prevention, and/or treatment of urinary system diseases. The carrier protein has excellent transmucosal efficiency, can realize transmucosal administration through pulmonary administration and gastrointestinal administration, has long-acting and slow-release characteristics in vivo, and realizes transmucosal drug delivery through fusion expression, chemical grafting and other modes by carrying functional molecules for diagnosing, preventing and/or treating the urinary system diseases, thereby realizing targeted diagnosis and treatment of the urinary system diseases.

Inventors

  • TIAN RUI
  • XIE SHIQIANG
  • HAN DANDAN
  • FENG XIN
  • Yang shengren
  • XIA NINGSHAO

Assignees

  • 翔安创新实验室
  • 厦门大学

Dates

Publication Date
20260505
Application Date
20251031
Priority Date
20241031

Claims (20)

  1. 1. A complex comprising a carrier protein and a functional molecule for the diagnosis, prevention and/or treatment of a urinary system disorder; Wherein the carrier protein is selected from the group consisting of a mutant DIIIbV of the subfraction of the albumin third domain (e.g., DIIIb), an albumin third domain DIIIV containing the mutant, and a multimer of DIIIbV or DIIIV.
  2. 2. The complex of claim 1, wherein the mutant comprises an insertion, substitution, deletion, and/or mutation of 1 or more (e.g., 40-50, 30-40, 20-30, 15-20, 10-15, 5-10, 1-5) amino acids compared to the wild type; preferably, the inserted, substituted, deleted and/or mutated amino acids correspond to the amino acids at positions 467 to 585 of SEQ ID NO. 1; More preferably, the inserted, substituted, deleted and/or mutated amino acids correspond to the amino acids at positions 497 to 585 of SEQ ID NO. 1; More preferably, the inserted, substituted, deleted and/or mutated amino acids correspond to the amino acids at positions 500 to 573 of SEQ ID NO. 1.
  3. 3. The complex of claim 1 or 2, said DIIIbV or DIIIV: (1) Having a higher affinity for FcRn under acidic or weakly acidic conditions as compared to wild-type, preferably the DIIIV affinity Kd values for FcRn are at the 10 -6 M,10 -7 M,10 -8 M,10 -9 M,10 -10 or 10 -11 M level, preferably the DIIIbV affinity Kd values for FcRn are at the 10 -6 M,10 -7 M,10 -8 M,10 -9 M,10 -10 or 10 -11 M level; (2) Mutations comprising 1,2, 3,4, 5,6, 7,8, 9, 10, 11, 12, 13, 14, 15 or more amino acids compared to the wild type; (3) A mutation comprising one or more amino acid positions selected from the group consisting of amino acids 500, 505, 523, 524, 527, 528, 531, 547, 509, 510, 498, 512 and 573 corresponding to a natural albumin having the amino acid sequence as set forth in SEQ ID NO. 1; (4) The mutation is a conservative mutation; (5) The part of the DIII comprises a fragment of natural albumin corresponding to amino acids 467 to 585 of SEQ ID No. 1; preferably, the DIIIbV or DIIIV comprises amino acid mutations at positions 523 and 573; Preferably, the DIIIbV or DIIIV comprises the amino acid mutations at positions 505, 523, 547 and 573; Preferably, the DIIIbV or DIIIV comprises amino acid mutations at positions 500, 505, 523, 524, 527, 528, 531, 547 and 573, or (6) Any combination of the above.
  4. 4. The complex of any one of claims 1-3, said DIIIbV or DIIIV: (1) The 500 th amino acid of the natural albumin with the amino acid sequence shown in SEQ ID NO. 1 is L or D; (2) At amino acid position 505 corresponding to natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is Q, N or T; (3) Amino acid 523, which corresponds to a natural albumin having the amino acid sequence shown in SEQ ID NO. 1, is L or M; (4) The amino acid at position 524 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is L; (5) The amino acid at position 527 which corresponds to the natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is K; (6) The amino acid at position 528 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is H or Y; (7) The amino acid at position 531 corresponding to natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is L; (8) The amino acid at position 547 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO.1 is A or C; (9) The amino acid at position 509 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO.1 is L; (10) The amino acid at position 510 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO.1 is R or N; (11) The amino acid at position 498 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is E; (12) The amino acid at position 512 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO.1 is G; (13) The amino acid at position 573 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO. 1 is any amino acid other than K (e.g., P), or (14) The amino acid at position 508 corresponding to the natural albumin having the amino acid sequence shown in SEQ ID NO.1 is L; (15) Any combination of the above.
  5. 5. The complex of claim 4, said DIIIbV or DIIIV: (1) The amino acid at position 500 is mutated from K to L or D; (2) The amino acid at position 505 is mutated from E to Q, N or T; (3) The amino acid at position 523 is mutated from I to L or M; (4) The amino acid at position 524 is mutated from K to L; (5) The amino acid at position 527 is mutated from T to K; (6) The amino acid at position 528 is mutated from a to H or Y; (7) Mutating the amino acid at position 531 from E to L; (8) The amino acid at position 547 is mutated from V to a or C; (9) The amino acid at position 509 is mutated from F to L; (10) The amino acid at position 510 is mutated from H to R or N; (11) The amino acid at position 498 is mutated from V to E; (12) Mutating the amino acid at position 512 from D to G; (13) The amino acid at position 573 is mutated from K to any amino acid other than K (e.g., P); (14) The amino acid at position 508 is mutated from T to L, or (15) Any combination of the above; Preferably, the DIIIbV or DIIIV comprises mutations I523G and K573P; Preferably, the DIIIbV or DIIIV comprises mutations E505Q, I523G, V547A and K573P; Preferably, the DIIIbV or DIIIV comprises the mutations K500L, E505Q, I523L, K524L, T527K, a528H, E531L, V547A and K573P.
  6. 6. The complex of any one of claim 1 to 5, wherein, Preferably, the mammal is selected from the group consisting of a human, a chimpanzee, a gorilla, a rhesus monkey, a rabbit, a mouse, a rat, a hamster, a cow, a horse, a donkey, a goat, a sheep, a dog, a guinea pig and a pig; preferably, the natural human serum albumin comprises or consists of the amino acid sequence shown in SEQ ID NO. 1; Preferably, the natural human serum albumin DIII comprises or consists of the amino acid sequence shown in SEQ ID No. 2; Preferably, the native human serum albumin DIIIb comprises or consists of the amino acid sequence shown in SEQ ID No. 4.
  7. 7. The complex of any one of claims 1-6, wherein DIIIbV is selected from the group consisting of the amino acid sequences shown in SEQ ID NOs 36-40; preferably, DIIIV is selected from the amino acid sequences shown in SEQ ID NO 5-35; Preferably, DIIIV has the amino acid sequence shown in SEQ ID NO. 5.
  8. 8. The complex of any one of claims 1-7, the carrier protein further comprising an albumin first domain DI, part, derivative or mutant thereof, and/or an albumin second domain DII, part, derivative or mutant thereof; Preferably, the DI and the DII are each independently derived from serum albumin native to a mammal, preferably the mammal is selected from the group consisting of a human, a chimpanzee, a gorilla, a rhesus monkey, a rabbit, a mouse, a rat, a hamster, a cow, a horse, a donkey, a goat, a sheep, a dog, a guinea pig and a pig, preferably the mammal is a human.
  9. 9. The complex of any one of claims 1-8, wherein the multimer is a homo-or hetero-dimer, trimer, tetramer, or any multimer that can be stably present in a physiological solution of the DIIIbV or DIIIV.
  10. 10. The complex of any one of claims 1-9, wherein the urinary system disease is kidney injury, urinary system inflammation, or urinary system cancer; preferably, the urinary system inflammation is selected from allergic inflammation such as glomerulonephritis, infections of the urinary tract such as tuberculosis of the kidney, pyelonephritis, cystitis, urethritis; preferably, the urinary system tumor is selected from renal cancer, urothelial cancer such as renal pelvis cancer, ureter cancer, bladder cancer, urethra cancer. Preferably, the kidney injury is selected from acute kidney injury, acute kidney disease, chronic kidney disease, or complications of other chronic and autoimmune diseases (e.g., diabetic nephropathy, systemic lupus erythematosus kidney disease, igA kidney disease); Preferably, the urinary system disease is selected from kidney disease, ureter disease, bladder disease and urinary tract disease; Preferably, the kidney disease is selected from the group consisting of kidney injury (e.g., acute kidney injury), glomerular disease, tubular-interstitial disease and kidney cancer, preferably the glomerular disease is selected from the group consisting of nephritis syndrome, nephrotic syndrome and diabetes-induced glomerular lesions (e.g., diabetic nephropathy), preferably the tubular-interstitial disease is selected from the group consisting of tubular-interstitial nephritis (e.g., acute or chronic tubular-interstitial nephritis), acute pyelonephritis, acute tubular necrosis, acute tubular nipple necrosis, obstructive or reflux nephropathy, nephrocaluria, nephropysis, kidney or perirenal abscess.
  11. 11. The complex of any one of claims 1-10, wherein the functional molecule is selected from a diagnostic agent, an imaging agent, a therapeutic radionuclide, or a large or small molecule drug, such as a polypeptide, a protein, an antibody, a nanobody, a nucleic acid drug, or a chemotherapeutic drug, preferably wherein the polypeptide or protein is a peptide chain or a cyclic peptide, preferably wherein the antibody is a monoclonal antibody or antigen-binding fragment thereof, and wherein the nucleic acid drug is an mRNA or protein nucleic acid complex; Preferably, the drug is an anti-inflammatory, anti-apoptotic or anti-oxidative stress drug against the urinary system disease, preferably, the drug is selected from amifostine, cimetidine, glutathione, RNLS agonists (e.g. RP 81), cortisone, curcumin, dexamethasone, and MMAE; Preferably, the diagnostic agent is selected from the group consisting of high-expression or high-secretion substances of the kidney system (e.g., interleukins, endogenous factor-vitamin B12 receptor (cubilin), low-density lipoprotein receptor-related protein 2 (megalin), kidney injury-related molecule (KIM-1), neutrophil gelatinase-related lipocalin (NGAL), and other proteins; Preferably, the imaging agent is selected from cyanine dyes such as IR-780, IR-783, and other photosensitizers; Preferably, the therapeutic radionuclide is selected from 64 Cu、 18 F、 68 Ga、 177 Lu、 125 I、 90 Y、 89 Sr、 32 P、 233 Ra.
  12. 12. The complex of any one of claims 1-11, wherein the carrier protein and the functional molecule are linked directly or through a linker; Preferably, the linker is a cleavable or non-cleavable linker, preferably the cleavable linker is of the acid cleavage type, disulfide cleavage type, protease cleavage type, glycosidase cleavage type or phosphatase cleavage type, preferably the linker is selected from DBCO-NHS ester、Sulfo-SMCC sodium、CL2 linker、DSP Cross linker、Mc-Val-Cit-PABC-PNP、Val-Cit-PAB、MC-Val-Cit-PAB、MAC glucuronide linker-2、Fmoc-PEA、Mal-PEG4-OH、3-Mercaptopropionic acid NHS ester and tBoc-NH-PEG-NH 2 .
  13. 13. The complex of any one of claims 1-12, wherein the carrier protein and the functional molecule are coupled to form the complex by genetic fusion or chemical means.
  14. 14. A nucleic acid molecule encoding the complex of any one of claims 1-13; preferably, the complex is a fusion protein.
  15. 15. A vector comprising the nucleic acid molecule of claim 14, preferably the vector is an expression vector; preferably, the vector is a eukaryotic bacterial vector (e.g., pPIC9K, pcdna 3.4).
  16. 16. A host cell comprising the nucleic acid molecule of claim 14 or the vector of claim 15; preferably, the cell is a eukaryotic cell or a prokaryotic cell; Preferably, the eukaryotic cells are yeast cells (e.g., saccharomyces cerevisiae, pichia pastoris) and 293T cells; Preferably, the prokaryotic cell is an E.coli cell, a B.subtilis cell, or any combination thereof.
  17. 17. A delivery combination or pharmaceutical composition comprising the complex of any one of claims 1-13; Preferably, the delivery combination or pharmaceutical composition is delivered intravenously; Preferably, the delivery combination or pharmaceutical composition is delivered via nasal or oral inhalation, preferably via nasal delivery, e.g. nasal drops, nasal spray or a combination thereof; Preferably, the complex is delivered to a mucosal surface of a subject (e.g., oral mucosa, nasal mucosa, tracheal mucosa, eyelid mucosa, vaginal mucosa); Preferably, the delivery combination or pharmaceutical composition further comprises one or more mucoadhesives to enhance the residence time of the effector molecule at the mucosal surface of the subject; Preferably, the delivery combination or pharmaceutical composition is an aerosol, powder spray, spray or other dosage form suitable for administration by inhalation; Preferably, the pharmaceutical composition contains one or more pharmaceutically acceptable auxiliary materials.
  18. 18. Use of the complex of any one of claims 1-13, the delivery combination of claim 17, or the pharmaceutical composition in the manufacture of a medicament for the treatment of a urinary system disorder; preferably, the urinary system disease is kidney injury, urinary system inflammation or urinary system cancer; preferably, the urinary system inflammation is selected from allergic inflammation such as glomerulonephritis, infections of the urinary tract such as tuberculosis of the kidney, pyelonephritis, cystitis, urethritis; preferably, the urinary system tumor is selected from renal cancer, urothelial cancer such as renal pelvis cancer, ureter cancer, bladder cancer, urethra cancer. Preferably, the kidney injury is selected from acute kidney injury, acute kidney disease, chronic kidney disease, or complications of other chronic and autoimmune diseases (e.g., diabetic nephropathy, systemic lupus erythematosus kidney disease, igA kidney disease); Preferably, the kidney disease is selected from the group consisting of kidney injury (e.g., acute kidney injury), glomerular disease, tubular-interstitial disease, and kidney cancer. In some embodiments, the glomerular disease is selected from the group consisting of nephritis, nephrotic syndrome, and diabetes-induced glomerular disease (e.g., diabetic nephropathy). In some embodiments, the tubular-interstitial disease is selected from tubular-interstitial nephritis (e.g., acute or chronic tubular-interstitial nephritis), acute pyelonephritis, acute tubular necrosis, acute papillary necrosis, obstruction or reflux nephropathy, nephrocalcareous pigmentation, nephropysis, kidney or perirenal abscess.
  19. 19. The complex of any one of claims 1-13, the delivery combination of claim 17, or the pharmaceutical composition for use in the treatment of a urinary system disorder; preferably, the urinary system disease is kidney injury, urinary system inflammation or urinary system cancer; preferably, the urinary system inflammation is selected from allergic inflammation such as glomerulonephritis, infections of the urinary tract such as tuberculosis of the kidney, pyelonephritis, cystitis, urethritis; preferably, the urinary system tumor is selected from renal cancer, urothelial cancer such as renal pelvis cancer, ureter cancer, bladder cancer, urethra cancer. Preferably, the kidney injury is selected from acute kidney injury, acute kidney disease, chronic kidney disease, or complications of other chronic and autoimmune diseases (e.g., diabetic nephropathy, systemic lupus erythematosus kidney disease, igA kidney disease); Preferably, the kidney disease is selected from the group consisting of kidney injury (e.g., acute kidney injury), glomerular disease, tubular-interstitial disease, and kidney cancer. In some embodiments, the glomerular disease is selected from the group consisting of nephritis, nephrotic syndrome, and diabetes-induced glomerular disease (e.g., diabetic nephropathy). In some embodiments, the tubular-interstitial disease is selected from tubular-interstitial nephritis (e.g., acute or chronic tubular-interstitial nephritis), acute pyelonephritis, acute tubular necrosis, acute papillary necrosis, obstruction or reflux nephropathy, nephrocalcareous pigmentation, nephropysis, kidney or perirenal abscess.
  20. 20. A method of treating a urinary system disorder comprising the step of administering to a subject in need thereof an effective amount of the complex of any one of claims 1-13, the delivery combination of claim 17, or the pharmaceutical composition; preferably, the urinary system disease is kidney injury, urinary system inflammation or urinary system cancer; preferably, the urinary system inflammation is selected from allergic inflammation such as glomerulonephritis, infections of the urinary tract such as tuberculosis of the kidney, pyelonephritis, cystitis, urethritis; preferably, the urinary system tumor is selected from renal cancer, urothelial cancer such as renal pelvis cancer, ureter cancer, bladder cancer, urethra cancer. Preferably, the kidney injury is selected from acute kidney injury, acute kidney disease, chronic kidney disease, or complications of other chronic and autoimmune diseases (e.g., diabetic nephropathy, systemic lupus erythematosus kidney disease, igA kidney disease); Preferably, the kidney disease is selected from the group consisting of kidney injury (e.g., acute kidney injury), glomerular disease, tubular-interstitial disease, and kidney cancer. In some embodiments, the glomerular disease is selected from the group consisting of nephritis, nephrotic syndrome, and diabetes-induced glomerular disease (e.g., diabetic nephropathy). In some embodiments, the tubular-interstitial disease is selected from tubular-interstitial nephritis (e.g., acute or chronic tubular-interstitial nephritis), acute pyelonephritis, acute tubular necrosis, acute papillary necrosis, obstruction or reflux nephropathy, nephrocalcareous pigmentation, nephropysis, kidney or perirenal abscess.

Description

Medicine for treating urinary system diseases The present application is based on and claims priority from chinese patent application number 202411547095.9, day 2024, month 10, 31, the disclosure of which is incorporated herein by reference in its entirety. Technical Field The application relates to the field of biological agents, in particular to a medicament for treating urinary system diseases. Background The mucous membrane is used as the first defense line of organism immune system and is widely distributed in the parts of oral cavity, nasal cavity, eye, gastrointestinal tract, vagina, respiratory tract, etc. It is a membranous structure composed of epithelial tissue and connective tissue, the epithelial portion of which is called epithelium, inside which there are rich blood vessels and nerves, covered with mucus. The mucosa drug delivery system is a novel drug delivery system, which plays a role in local action or absorption into systemic circulation to trigger systemic therapeutic action by applying drugs and proper carrier materials to mucosa parts. The mucous membrane administration has the advantages that firstly, compared with oral administration, the mucous membrane absorption of the medicine enters the systemic circulation, the medicine does not pass through the liver, the first pass effect of the liver can be effectively avoided, secondly, enzyme types and quantity in the mucous membrane are limited, the enzymatic degradation reaction activity of the medicine is relatively low, the stability of the medicine is improved, in addition, the local administration improves the medicine concentration of an acting part, further improves the bioavailability, and finally, the local administration enables the administration process to be more convenient, the patient can operate independently, and the compliance of the patient is improved. These advantages make mucosal administration have a broad application prospect in drug delivery. Kidneys are key organs for maintaining organism homeostasis, and play important physiological functions of filtering blood, removing metabolic waste, regulating water-salt balance and electrolyte, secreting hormone and the like. The structural and functional complexity determines the challenges and specificity of drug targeting. The physiological properties of the kidneys themselves present a technical hurdle for drug delivery. For a long time, kidney targeted drug delivery has two major difficulties, namely, kidney is taken as an important metabolic organ of a human body, after the drug enters blood, the drug is easily and rapidly excreted by the kidney, so that the drug is removed without reaching the effective drug concentration, the drug needs to be administered at high frequency to maintain the local effective drug concentration, the drug administration burden of a patient is increased, the adverse reaction risk of the drug is possibly increased, and the exogenous drug is difficult to accurately penetrate and concentrate in key lesion areas such as renal tubules and the like due to the complex hierarchical structure and physiological barriers (such as glomerular filtration barriers, renal tubular epithelial cell barriers and the like) of the kidney, so that the drug utilization rate is low and the treatment effect is limited. There is a need for a warhead that can achieve effective kidney targeting to solve the existing kidney disease dilemma. Disclosure of Invention It has been unexpectedly found that a protein with neonatal Fc receptor (neonatal Fc receptor, fcRn) affinity can efficiently deliver a variety of loads (small molecule drugs, proteins, nucleic acids, etc.), and can be reabsorbed into blood via proximal renal tubules by administration through a mucosal system or by administration through blood, thereby realizing long-term renal targeting. According to different requirements, nasal spray administration, bronchial administration, oral mucosa, gastrointestinal mucosa, blood or other modes can be selected for administration, and a brand new tool is provided for accurate diagnosis, long-term monitoring and efficient treatment of kidney diseases. Neonatal Fc receptor (neonatal Fc receptor, fcRn) is a receptor for human serum albumin (Human serum albumin, HSA), mediating lysosomal escape of HSA to achieve long circulation of albumin in vivo. The protein structure of FcRn is a heterodimer composed of a non-covalent association of an alpha chain and beta 2 microglobulin, is widely expressed in various tissue epithelial cells, endothelial cells and various immune cells in vivo, and FcRn is highly expressed on the surface of mucosal epithelial cells (e.g., nasal airway and olfactory epithelium). FcRn high affinity albumin DIII mutants were screened as kidney targeting warheads by constructing FcRn different affinity size (high affinity or low affinity) mutants and high throughput screening methods. The high affinity mutant has excellent transmucosal efficiency and lon