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CN-122012498-A - SiRNA for inhibiting INHBE gene expression, conjugate and application thereof

CN122012498ACN 122012498 ACN122012498 ACN 122012498ACN-122012498-A

Abstract

The invention belongs to the field of biological medicine, and in particular relates to siRNA for inhibiting INHBE gene expression, a conjugate and application thereof. The invention designs corresponding siRNA aiming at INHBE genes, delivers the siRNA to the liver through coupling GalNAc, and interferes INHBE mRNA at the liver part, thereby effectively reducing INHBE protein expression, promoting healthy fat storage and further playing a role in treating obesity. The siRNA and the siRNA conjugate provided by the invention can obviously inhibit proliferation of human hepatoma cell Hep-G2 and have an effect on the inhibition rate of mouse liver INHBE mRNA, so that the siRNA and the siRNA conjugate provided by the invention have good prospects in application of obesity.

Inventors

  • ZHANG YU
  • ZHAO CHENGJIANG
  • HUANG JINGJING
  • ZHANG HANXIN

Assignees

  • 佑嘉(杭州)生物医药科技有限公司

Dates

Publication Date
20260512
Application Date
20251110
Priority Date
20241111

Claims (13)

  1. 1. The siRNA for inhibiting INHBE gene expression is characterized by comprising a sense strand and an antisense strand, wherein the sense strand and the antisense strand are at least partially reversely complementary to form a double-stranded region, the sense strand of the siRNA has a nucleotide sequence shown as SEQ ID NO. 1-11, and the antisense strand has a nucleotide sequence shown as SEQ ID NO. 13-23.
  2. 2. The siRNA of claim 1, wherein at least one nucleotide in said sense strand or said antisense strand is a modified nucleotide.
  3. 3. The siRNA of claim 2 wherein at least one nucleotide in said sense strand or said antisense strand is a nucleotide analog comprising an iso-nucleotide, LNA, ENA, cEt-BNA, UNA, or GNA or a hydroxy-modified nucleotide at the 2' -position of the ribosyl, said hydroxy-modified nucleotide at the 2' -position of the ribosyl comprising a 2' -O-methyl, 2' -O-methoxyethyl, 2' -O-aminopropyl, 2' -deoxy, T-deoxy-2 ' -fluoro, 2' -O-aminopropyl, 2' -O-dimethylaminoethyl, 2' -O-dimethylaminopropyl, T-O-dimethylaminoethoxyethyl, or 2' -O-N-methylacetamido-modified nucleotide.
  4. 4. The siRNA of claim 2, wherein at least one phosphate group in the sense strand or the antisense strand is a phosphate group having a modifying group, and wherein the phosphate group is a phosphorothioate group formed by substitution of at least one oxygen atom in a phosphodiester bond with a sulfur atom.
  5. 5. The siRNA of any one of claims 2 to 4, wherein the sense strand structure of the modified siRNA molecule is shown in SEQ ID No. 25, the antisense strand structure of the modified siRNA molecule is shown in SEQ ID No. 27, or the antisense strand structure of the modified siRNA molecule is shown in SEQ ID No. 28, the antisense strand structure of the modified siRNA molecule is shown in SEQ ID No. 30, or the antisense strand structure of the modified siRNA molecule is shown in SEQ ID No. 57, and the antisense strand structure of the modified siRNA molecule is shown in SEQ ID No. 59.
  6. 6. An siRNA conjugate comprising the siRNA of any one of claims 1 to 5 and a ligand conjugated to the siRNA.
  7. 7. The siRNA conjugate of claim 6, wherein the ligand comprises N-acetylgalactosamine, an aliphatic, a cycloaliphatic, a polycycloaliphatic compound, cholesterol, biotin, a vitamin, galactose, lactose, N-acetylglucosamine, or a derivative thereof.
  8. 8. The siRNA conjugate of claim 7, wherein the ligand is GalNAc or a derivative thereof.
  9. 9. The siRNA conjugate of claim 8, the ligand being one or more GalNAc derivatives attached by a monovalent, divalent or trivalent branching linker.
  10. 10. The siRNA conjugate of claim 9, wherein the ligand is L96, and the L96 is N- (tris (GalNAc-alkyl) -amide decanoyl) -4-hydroxy prolyl.
  11. 11. The biomaterial related to the siRNA of any one of claims 1 to 10, being any one of the following: 1) A vector comprising the siRNA of any one of claims 1 to 10; 2) A reagent or kit comprising the siRNA of any one of claims 1 to 10 or the vector of 1); 3) A pharmaceutical composition comprising the siRNA molecule of any one of claims 1 to 10 and a pharmaceutically acceptable additional component.
  12. 12. Use of the siRNA of claims 1-5 or the siRNA conjugate of claims 6-10 or the pharmaceutical composition of claim 11 for the preparation of a medicament for alleviating and/or treating obesity.
  13. 13. Use of the siRNA of claims 1-5 or the siRNA conjugate of claims 6-10 or the pharmaceutical composition of claim 11 in combination with semaglutin.

Description

SiRNA for inhibiting INHBE gene expression, conjugate and application thereof Technical Field The invention relates to the field of biological medicine, in particular to siRNA for inhibiting INHBE gene expression, a conjugate and application thereof. Background RNA interference (RNA INTERFERENCE, abbreviated RNAi) refers to a phenomenon of gene silencing induced by double-stranded RNA in molecular biology by inhibiting gene expression by blocking transcription or translation of a particular gene. When double-stranded RNA homologous to the coding region of an endogenous messenger RNA (MESSENGER RNA, abbreviated mRNA) is introduced into a cell, the mRNA is degraded and gene expression is silenced. The small interfering RNA (SMALL INTERFERING RNA, SIRNA) with the length of 20-25nt can trigger RNAi, can specifically down regulate or close the expression of specific genes, has the characteristics of high efficiency, easy synthesis, easy operation and the like, so the technology has been widely used in the field of exploring gene functions and gene therapy of infectious diseases and malignant tumors. Acetylgalactosamine (GalNAc) is a monosaccharide that recognizes with sialoglycoprotein receptors that are highly expressed by hepatic parenchymal cells. The use of derivatives of GalNAc, such as bivalent or trivalent branched linkers, attached to the 3' end of the sense strand of the siRNA can promote specific targeting of the siRNA to liver tissue, thereby improving its bioavailability and reducing the dose and side effects of administration. Obesity has shown a rapidly growing situation worldwide over the last decades. World Health Organization (WHO) data showed that the number of obese people worldwide has increased nearly twice since 1975. In 2016, over 19 million adults (18 years and older) were overweight worldwide, with over 6.5 million obese. The prevalence of obesity in high-income countries is relatively high, but in recent years, the prevalence of obesity in low-and medium-income countries has also increased dramatically. For example, in north america, the united states is one of the countries where obesity problems are more serious, with adult obesity prevalence exceeding 30%. In some Pacific islands, such as Nauru and Cookry islands, the obesity rate is more high up to 70% -90%. In the middle east and northern africa, the obesity rate is also continuously rising, and the obesity problem of countries like Egypt is increasingly prominent. In asia, although the obesity rate is traditionally low, the number of obese people is rapidly increasing with economic development and lifestyle changes. For example, china and india, the absolute number of obese people increases significantly due to the huge population base. The obesity rate of China is increased from the past lower level to about 10% -15% of the obesity rate of adults in the urban area. At the physiological level, obesity is caused by the long term energy intake exceeding energy expenditure, and the excess energy is stored in the body in the form of fat. This process involves an imbalance in complex metabolic regulation mechanisms, including multiple factors such as neuroendocrine system, abnormal secretion of adipocytes, and disturbances in intestinal flora. For example, the hypothalamic-pituitary-adrenal axis and hypothalamic-pituitary-thyroid axis in the neuroendocrine system play a key role in the regulation of energy balance, once unbalanced, affecting appetite and metabolism. Whereas alterations in the secretion of adipocytes such as leptin, adiponectin, etc. interfere with insulin sensitivity and energy metabolism. Intestinal flora is also closely related to obesity, and dysbacteriosis may affect food digestion and absorption and energy harvesting. Obesity is closely connected with various serious diseases, is an important risk factor of cardiovascular diseases, type 2 diabetes, hypertension, sleep apnea hypoventilation syndrome, nonalcoholic fatty liver disease, osteoarthritis, various cancers and other diseases, brings heavy medical burden to patients, and also causes great pressure on social medical resources. Existing therapies for obesity mainly include lifestyle interventions, drug therapies and surgical therapies. Diet control in lifestyle interventions is to reduce weight by adjusting dietary structure, controlling caloric intake, such as increasing dietary fiber food intake, reducing "three-high" food intake and controlling meal size and total calories, but it relies on patient's own discipline, with great difficulty in adherence and slow effect on severely obese people. Exercise includes aerobic and strength training to increase energy consumption, can be limited by physical conditions, time and environment, and is not easy to develop regular exercise habits because the movement of obese people is easy to damage joints. In the aspect of drug treatment, appetite suppressants such as phentermine can stimulate hypothalamic satiety center