WO2021047607A1 - 非igf1r结合型的物质在预防和/或治疗炎症性疾病中的应用 - Google Patents
非igf1r结合型的物质在预防和/或治疗炎症性疾病中的应用 Download PDFInfo
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Definitions
- the present invention relates to the field of biomedicine, in particular to the application of non-IGF1R-binding substances in the prevention and/or treatment of inflammatory diseases.
- IGF2 Insulin like growth factor 2
- IGF2 receptors include IGF1R and IGF2R, among which IGF2R has the highest affinity, followed by binding to IGF1R, and almost no binding to insulin receptor 2 .
- IGF2 tyrosine kinase IGF1R open, but with no positive correlation IGF2R and IR 2.
- IGF2R Intracellular Golgi apparatus
- IGF2R is not the only receptor of IGF2
- IGF2 is not the only ligand of IGF2R. Its ligands also include mannose-6-phosphate.
- the main function of IGF2R is to transport its ligands to lysosomes. 4 complete protein processing or degradation.
- studies have found that, the IGF2R on the cell membrane can be cut off in a soluble form, into the blood and other liquid environment, effective binding IGF2, 4,5 inhibition IGF2 function.
- Autoimmune diseases are diseases caused by the immune system's abnormal attacks on one's own organs and tissue cells. According to data released by the US Department of Health and Human Services, there are currently 24 million Americans-7% of the US population suffers from more than 80 types. The torment of autoimmune diseases involves almost every part of the body6,7 . The pathogenic factors of most autoimmune diseases are not yet clear, and may be related to factors such as genetics, infections, drugs, and the environment. The disease course is longer, with repeated remissions and attacks, and more women than men are affected. Most autoimmune diseases involve the participation of phagocytes composed of monocyte precursor cells, monocytes and macrophages.
- activated macrophages secrete milk fat globule-epidermal growth factor 8 (MFG-E8), the C-terminal domain of MFG-E8 and apoptotic cells phosphatidylserine binding, mediates phagocytosis of apoptotic cells, autoimmune diseases clear attack and death in apoptosis and immune cell debris, to help the body to restore homeostasis 8.
- MFG-E8 milk fat globule-epidermal growth factor 8
- apoptotic cells phosphatidylserine binding
- MFG-E8-mediated macrophage phagocytosis of apoptotic fragments can also inhibit the regulation of lipopolysaccharide and other pro-inflammatory effects on macrophages, including p38, ERK1/2, c-JNK, MAPK and 10 inhibition of p65.
- Inflammation is the body's defense response to its own and foreign inflammatory factors. Inflammatory factors include biological factors, physical factors, chemical factors, foreign bodies, necrotic tissues, allergic reactions and pathological changes. Inflammatory reactions are affected by inflammatory factors. 11 and the body's own regulation. In inflammatory diseases in both natural immunity to participate, but also to participate in adaptive immunity, in the early innate immune generated inflammation lead to tissue injury site, and determine the impact of late innate and adaptive immune response type 12. When the inflammatory regulation is disordered or obstructed, inflammatory diseases occur, and the ability to resist stimuli and pathogens is lost, which accelerates the aggravation of the disease. According to the speed of the inflammatory response, the inflammatory response includes both acute and chronic inflammation.
- the first category is steroidal anti-inflammatory drugs, that is, steroid drugs, such as adrenal cortex hormones, male hormones, and estrogen, which have certain anti-inflammatory effects. long-term use will cause endocrine disorders such as multi-system dysfunction 13-15.
- the second category is non-steroidal anti-inflammatory drugs including aspirin, acetaminophen, indomethacin, naproxen, naproxen, diclofenac, ibuprofen, nimesulide, rofecoxib, celecoxib etc., it is widely used in clinical osteoarthritis, rheumatoid arthritis and other anti-inflammatory treatment 14-16.
- Nonsteroidal anti-inflammatory drug single action principle primarily anti-inflammatory action by inhibiting the synthesis of prostaglandin 17, and reduce the formation of bradykinin, and reduce platelet aggregation and leukocyte aggregation. Therefore, sterol drugs are limited by side effects, while non-steroid drugs are limited by a single pathway of action, and the combination of drugs has a poor sensitization effect.
- the purpose of the present invention is to provide a medicine that can treat autoimmune diseases and/or inflammatory diseases more effectively.
- the first aspect of the present invention provides the use of a non-IGF1R-binding substance for the preparation of a composition or preparation for the prevention and/or treatment of inflammatory diseases.
- the non-IGF1R-binding substance is selected from the group consisting of IGF2 mutants, vectors expressing IGF2 mutants, antibodies, small molecule compounds, or combinations thereof.
- the inflammatory disease is selected from the group consisting of peritonitis, inflammatory bowel disease, multiple sclerosis, diabetes, systemic lupus erythematosus, scleroderma, Hashimoto's thyroiditis, autoimmune hepatitis, self Immune uveitis, interstitial lung disease, psoriasis, vitiligo, dermatomyositis, Kawasaki disease, adult ear disease, compulsive spondylitis, sarcoidosis, arthritis associated with start and end inflammation, polyarticular adolescents Idiopathic arthritis, rheumatoid arthritis, graft-versus-host disease, autoimmune pancreatitis, Parkinson's disease, Alzheimer's disease, or a combination thereof.
- the non-IGF1R-binding substances include substances that have a higher selectivity or affinity for IGF2R than IGF1R, and substances that can efficiently activate IGF2R that do not activate or substantially do not activate IGF1R (such as retinoic acid, etc.).
- Non-IGF1R ligand Non-IGF1R ligand
- the vector expressing the IGF2 mutant includes a viral vector.
- the viral vector is selected from the following group: adenovirus vector, lentivirus vector, or a combination thereof.
- the non-IGF1R-binding substance also includes one or more substances selected from the group consisting of Plasminogen, Serglycin, and e1a stimulation.
- Cellular suppressor of E1A-stimulated genes CREG
- Sgsh sulfamidase
- phosphorylated ⁇ -glucuronidase Phosphorylated ⁇ -glucuronidase, Gusb
- the IGF2 mutant has a mutation in the tyrosine corresponding to the 27th position of SEQ ID NO.:1 of the wild-type IGF2 protein.
- the tyrosine at position 27 is mutated into one or more amino acids selected from the group consisting of leucine, isoleucine, valine, methionine, and alanine. Acid, phenylalanine, serine, proline, threonine, histidine, lysine, tryptophan, arginine, glutamic acid, glycine, aspartic acid, cysteine.
- the tyrosine at position 27 is mutated to leucine.
- the IGF2 mutant mutates the tyrosine at position 27 of the wild-type IGF2 protein corresponding to SEQ ID NO.:1 to leucine, and optionally deletes D-domain (SEQ ID NO.:1). ID NO.:1 threonine 62 to glutamic acid 67).
- the IGF2 mutant has a mutation in the glutamic acid at position 12 of the wild-type IGF2 protein corresponding to SEQ ID NO. 1, and optionally deletes D-domain (SEQ ID NO. 1). :1 threonine at position 62 to glutamic acid at position 67).
- the glutamic acid at position 12 is mutated to one or more amino acids selected from the group consisting of Asp, Ala, Gln, His, Arg, and Lys.
- the IGF2 mutant has a mutation in the phenylalanine at position 26 of the wild-type IGF2 protein corresponding to SEQ ID NO. 1, and optionally deletes the D-domain (SEQ ID NO. .:1 threonine 62 to glutamic acid 67).
- the phenylalanine at position 26 is mutated to one or more amino acids selected from the group consisting of Ser, Asp, Ala, Gln, His, Arg, and Lys.
- the phenylalanine at position 26 is mutated to serine.
- the IGF2 mutant has a mutation in the tyrosine at position 27 of the wild-type IGF2 protein corresponding to SEQ ID NO. 1, and optionally deletes D-domain (SEQ ID NO. 1). :1 threonine 62 to glutamic acid 67).
- the tyrosine at position 27 is mutated to one or more amino acids selected from the group consisting of Leu, Asp, Ala, Gln, His, Arg, Lys.
- the tyrosine at position 27 is mutated to leucine.
- the IGF2 mutant has a mutation in the wild-type IGF2 protein corresponding to SEQ ID NO.:1, the 43rd valine, and optionally deletes D-domain (SEQ ID NO. 1). :1 threonine 62 to glutamic acid 67).
- the valine at position 43 is mutated to one or more amino acids selected from the group consisting of Leu, Asp, Ala, Gln, His, Arg, Lys.
- valine at position 43 is mutated to leucine.
- the tyrosine at position 27 is mutated to one or more amino acids selected from the group consisting of Leu, Asp, Ala, Gln, His, Arg, Lys, and the tyrosine at position 43 is Valine is mutated to one or more amino acids selected from the group consisting of Leu, Asp, Ala, Gln, His, Arg, Lys.
- the tyrosine at position 27 and the valine at position 43 are simultaneously mutated to leucine.
- the tyrosine at position 27 and the valine at position 43 are mutated to leucine at the same time, and the 62th threon of D-domain (SEQ ID NO.:1) is optionally deleted. Amino acid to glutamic acid at position 67).
- amino acid sequence of the IGF2 mutant is shown in any one of SEQ ID NO.: 2-58.
- the IGF2 mutant is a polypeptide having an amino acid sequence shown in any one of SEQ ID NO.: 2-58, an active fragment thereof, or a conservative variant polypeptide thereof.
- the IGF2 mutant except for the mutations (such as Glu at position 12, Phe at position 26, Tyr at position 27, Val at position 43 and the corresponding mutant sequence for deleting D-domain), the rest of the amino acid sequence It is the same or substantially the same as the sequence shown in SEQ ID NO.:1.
- the glutamic acid at position 12 is mutated to glutamine.
- the said substantially identical is at most 50 (preferably 1-20, more preferably 1-10, more preferably 1-5) amino acids are different, wherein, The difference includes the substitution, deletion or addition of amino acids (such as the deletion of T at position 62 to E at position 67), and the IGF2 mutant has the activity of inhibiting inflammation.
- the homology with the sequence shown in SEQ ID NO.:1 is at least 80%, preferably at least 85% or 90%, more preferably at least 95%, and most preferably at least 98% or 99%.
- the IGF2 protein is derived from a human or non-human mammal.
- the IGF2 mutant is selected from the following group:
- amino acid sequence shown in SEQ ID NO.: 2-58 is formed by the substitution, deletion or addition of one or more (such as 2, 3, 4 or 5) amino acid residues, It also has a polypeptide derived from (a) that inhibits inflammation.
- the homology between the derived polypeptide and any sequence shown in SEQ ID NO.: 2-58 is at least 60%, preferably at least 70%, and more preferably at least 80%. %, preferably at least 90%, such as 95%, 97%, 99%.
- the IGF2 mutant is formed by mutation of the wild-type IGF2 protein shown in SEQ ID NO.:1.
- composition or preparation is also used for one or more purposes selected from the following group:
- the autoimmune disease is selected from the group consisting of multiple sclerosis, inflammatory bowel disease, autoimmune hepatitis, systemic lupus erythematosus, rheumatoid arthritis, insulin resistance, diabetes, autoimmune Hepatitis, vitiligo, psoriasis, peritonitis, scleroderma, Hashimoto's thyroiditis, graft-versus-host disease, dermatomyositis, Kawasaki disease, adult ear disease, mandatory spondylitis, sarcoidosis, start and stop Spot inflammation-related arthritis, polyarticular juvenile idiopathic arthritis, autoimmune pancreatitis, and some undefined autoimmune diseases, or combinations thereof.
- the composition includes a pharmaceutical composition.
- the composition comprises (a) a non-IGF1R-binding substance; and (b) a pharmaceutically acceptable carrier.
- the composition contains 0.001-99wt%, preferably 0.1-90wt%, more preferably 1-80wt% of non-IGF1R-binding substances, based on the total weight of the composition.
- the composition is a liquid preparation or a lyophilized preparation.
- the composition is an injection.
- the second aspect of the present invention provides a cell preparation including:
- Phagocytes treated with non-IGF1R-binding substances Phagocytes treated with non-IGF1R-binding substances.
- the phagocytic cell is selected from the group consisting of monocytes, macrophages, monocyte precursor cells, or a combination thereof.
- the non-IGF1R-binding substance is selected from the following group: IGF2, IGF2 mutant, vector expressing IGF2 mutant, antibody, small molecule compound, or a combination thereof.
- the non-IGF1R-binding substance also includes one or more substances selected from the group consisting of Plasminogen, Serglycin, and e1a stimulation.
- Cellular suppressor of E1A-stimulated genes CREG
- Sulfamidase Sgsh
- phosphorylated ⁇ -glucuronidase Phosphorylated ⁇ -glucuronidase, Gusb
- the cell preparation further includes a non-IGF1R-binding substance.
- the phagocytic cell has characteristics selected from the following group:
- IGF1R is not activated or the degree of activation is lower than IGF2R, IGF2R is activated;
- Phagocytes use oxidative phosphorylation as the main way of obtaining energy; and/or
- the phagocytic cells are derived from bone marrow, abdominal cavity, peripheral blood, site of inflammation, or a combination thereof.
- the cell preparation includes a liquid preparation.
- the cells in the cell preparation basically ( ⁇ 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%) or all of them are passed through (a) Non-IGF1R-binding substance pretreated phagocytic cells and (b) optional non-IGF1R-binding substance constituted.
- the concentration of the phagocytic cells is 1 ⁇ 10 4 -5 ⁇ 10 7 /ml, preferably 5 ⁇ 10 4 -5 ⁇ 10 6 /ml, more preferably 1 ⁇ 10 5 -1 ⁇ 10 6 /ml.
- the carrier is selected from the following group: infusion carrier and/or injection carrier, preferably, the carrier is one or more carriers selected from the group: physiological saline, glucose Salt water, or a combination thereof.
- the cell preparation also includes other drugs for preventing and/or treating inflammatory diseases.
- the other drugs for preventing and/or treating inflammatory diseases are selected from the group consisting of non-steroidal anti-inflammatory drugs, glucocorticoids, methotrexate, TNF ⁇ neutralizing antibody, TNFR1 antibody, TNFR2 Antibodies, anti-CD20 antibodies, IL-1R antagonists, IL-12 and IL-23p40 neutralizing antibodies, IL-23p19 neutralizing antibodies, IL-17 neutralizing antibodies, IL-17A receptor neutralizing antibodies, or combinations thereof.
- the other drugs for preventing and/or treating autoimmune diseases are selected from the group consisting of non-steroidal anti-inflammatory drugs, glucocorticoids, methotrexate, TNF ⁇ neutralizing antibody, and TNFR1 antibody , TNFR2 antibody, anti-CD20 antibody, IL-1R antagonist, IL-12 and IL-23p40 neutralizing antibody, IL-23p19 neutralizing antibody, IL-17 neutralizing antibody, IL-17A receptor neutralizing antibody, CTLA- 4 Fusion protein, or a combination thereof.
- the third aspect of the present invention provides a medicine kit including:
- the kit further includes:
- the third container, and the active ingredient contained in the third container (c) other medicines for preventing and/or treating inflammatory diseases, or medicines containing the active ingredient (c).
- the kit further includes:
- the fourth container, and the active ingredient contained in the fourth container (d) Other drugs for preventing and/or treating autoimmune diseases, or drugs containing other drugs for preventing and/or treating autoimmune diseases .
- first container, the second container, the optional third container, and the optional fourth container may be the same or different.
- the medicine in the first container is a unilateral preparation containing phagocytes treated with a non-IGF1R-binding substance.
- the medicine in the second container is a unilateral preparation containing non-IGF1R-binding substances.
- the third container is a single preparation containing other drugs for preventing and/or treating inflammatory diseases.
- the fourth container is a single preparation containing other drugs for preventing and/or treating autoimmune diseases.
- the dosage form of the drug is an injection dosage form.
- the kit also contains instructions, which describe the combined administration of active ingredient (a), active ingredient (b), optional (c) and optional (d) so as to ( i) prevention and/or treatment of inflammatory diseases; and/or (ii) instructions for prevention and/or treatment of autoimmune diseases.
- the concentration of the non-IGF1R-binding substance is 0.01 ⁇ g-1 mg/kg body weight, preferably 0.1-10 ⁇ g/kg Body weight, more preferably 0.5-5 ⁇ g/kg body weight.
- the concentration of the phagocytic cells is 0.2*10 4 -1*10 8 cells/kg body weight, which is higher than Best 1*10 5 -5*10 7 /kg body weight, more preferably 1*10 6 -5*10 6 /kg body weight.
- the concentration of the other drugs for preventing and/or treating inflammatory diseases is 0.0002-40 mg/kg body weight , Preferably 0.001-30 mg/kg body weight, more preferably 0.02-25 mg/kg body weight.
- the concentration of the other drugs for preventing and/or treating autoimmune diseases is 0.0002 to 0.1 mg /kg body weight, preferably 0.001-0.08 mg/kg body weight, more preferably 0.002-0.01 mg/kg body weight.
- the fourth aspect of the present invention provides a drug screening method, including the steps:
- test group In the test group, add a test substance to the cell culture system, and observe the binding activity of the test substance with IGF1R and/or IGF2R in the cells of the test group; in the control group, the same cell culture No test substances are added to the system;
- the binding activity of the test substance to IGF1R in the cells of the test group decreases; and the binding activity to IGF2R remains unchanged or increases, it indicates that the test substance is a non-IGF1R-binding IGF2 mutant type.
- the method further includes the steps:
- step (b) For the non-IGF1R binding type IGF2 mutant obtained in step (a), further determine its binding activity to the IGFBP protein; and/or further test whether its binding to the IGFBP protein remains unchanged or slightly increased.
- the method further includes the steps:
- the positive control is IGF2 protein or IGF2 mutant, and its binding activity to IGF1R and/or IGF2R is compared with the non-IGF1R binding type IGF2 mutant obtained in step (a) to IGF1R and / Or the binding activity of IGF2R for comparison.
- the binding activity of the non-IGF1R-binding IGF2 mutant obtained in step (a) to IGF1R and/or IGF2R is equivalent to the binding activity of the positive control to IGF1R and/or IGF2R.
- a method for screening candidates for the prevention and/or treatment of inflammatory diseases including:
- test group In the test group, add a test substance to the cell culture system, and observe the effect of the test substance in the cells of the test group on the expression and/or activity of IGF2R on the cell surface; and/or the test The influence of the substance on the nuclear transport of IGF2R; and/or the influence of the test substance on the redistribution of IGF2R to the nucleus; in the control group, no test substance is added to the culture system of the same cell;
- test substance of the test group inhibits the expression and/or activity of IGF2R on the cell surface; and/or the test substance increases the nuclear transport of IGF2R; and/or the test substance promotes the redistribution of IGF2R to the nucleus, This indicates that the test substance is a candidate for the prevention and/or treatment of inflammatory diseases.
- the candidate has IGF2R selectivity; that is, the candidate has a greater selectivity for IGF2R than for IGF1R.
- the candidate selectively and/or preferentially binds to IGF2R, but does not bind or substantially does not bind to IGF1R.
- the cell is a cell cultured in vitro.
- the cell is a phagocytic cell.
- the phagocytic cell is selected from the group consisting of monocytes, macrophages, monocyte precursor cells, or a combination thereof.
- the method is an in vitro method.
- the method is non-diagnostic and non-therapeutic.
- the fifth aspect of the present invention provides a drug screening method (or a method for screening candidates), including:
- test group In the test group, add a test substance to the cell culture system, and observe the effect of the test substance on the binding activity of IGF2 and IGF1R and/or IGF2R in the cells of the test group; in the control group, No test substance is added to the same cell culture system;
- test substance in the cells of the test group inhibits the binding activity of IGF2 and IGF1R; and has no influence or promotion effect on the binding activity of IGF2 and IGF2R, it indicates that the test substance is a non-IGF1R-binding substance ( Or candidate).
- the non-IGF1R-binding substance is selected from the following group: antibodies, small molecule compounds, or a combination thereof.
- the method further includes the steps:
- step (b) For the non-IGF1R-binding substance obtained in step (a), further determine its binding activity to IGFBP protein; and/or further test whether its binding to IGFBP protein remains unchanged or slightly increases.
- the method further includes the steps:
- the positive control is IGF2 protein or IGF2 mutant, and its binding activity to IGF1R and/or IGF2R is compared with the non-IGF1R binding substance obtained in step (a) to IGF1R and/or The binding activity of IGF2R was compared.
- the binding activity of the non-IGF1R-binding substance obtained in step (a) to IGF1R and/or IGF2R is equivalent to the binding activity of the positive control to IGF1R and/or IGF2R.
- the cell is a cell cultured in vitro.
- the cell is a phagocytic cell.
- the phagocytic cell is selected from the group consisting of monocytes, macrophages, monocyte precursor cells, or a combination thereof.
- the method is an in vitro method.
- the method is non-diagnostic and non-therapeutic.
- the sixth aspect of the present invention provides a method for obtaining pro-inflammatory samples and anti-inflammatory samples in vitro, including:
- the binding activity of IGF2 to IGF2R is further determined.
- the binding activity of IGF2 to IGF2R is further determined.
- the seventh aspect of the present invention provides a composition comprising:
- non-IGF1R-binding IGF2 mutant or candidate obtained by the method of the fourth or fifth aspect of the present invention does not include an IGF2 mutation selected from the following group Type: wild-type IGF2, wild-type IGF2 amino acid fragments 25 to 91, IGF2 mutant with mutation of glutamic acid at position 12 to Asp, Ala, Gln, His, Arg, or Lys, mutation of phenylalanine at position 26 to Ser IGF2 mutant type, tyrosine 27 is mutated to Leu IGF2 mutant, valine 43 is mutated to Leu IGF2 mutant, IGF2 mutant with D-domain deleted, or a combination thereof.
- the non-IGF1R binding type IGF2 mutant does not include the IGF2 shown in any of SEQ ID NO.: 1-2, 16, 18, 20, 22, 24, 26, 28, 43 Mutant.
- amino acid sequence of the non-IGF1R-binding IGF2 mutant is shown in SEQ ID NO.: 3-15, 17, 19, 21, 23, 25, 27, 29-42, 44-58 Either shown.
- the eighth aspect of the present invention provides a method for inhibiting inflammatory activity, including the steps:
- phagocytic cells are cultured to suppress inflammatory activity.
- the application concentration of the non-IGF1R-binding substance is 0.0000005-0.0005 mg/ml, preferably, 0.000001-0.0001 mg/ml, more preferably, 0.000005-0.00005 mg/ml.
- the ninth aspect of the present invention provides a method for preventing and/or treating inflammatory diseases, including:
- the subject includes non-human mammals and humans.
- the dosage of the non-IGF1R-binding substance is 0.01 ⁇ g-1 mg/kg body weight, preferably 0.1-10 ⁇ g/kg body weight, more preferably 0.5-5 ⁇ g/kg body weight.
- the frequency of administration of the non-IGF1R-binding substance is 1 time/day to 2 times/day.
- the administration includes simultaneous administration or sequential administration.
- the tenth aspect of the present invention provides a method for preventing and/or treating autoimmune diseases, including:
- the subject includes non-human mammals and humans.
- the dosage of the non-IGF1R-binding substance is 0.01 ⁇ g-1 mg/kg body weight, preferably 0.1-10 ⁇ g/kg body weight, more preferably 0.5-5 ⁇ g/kg body weight.
- the frequency of administration of the non-IGF1R-binding substance is 1 time/day to 2 times/day.
- the administration includes simultaneous administration or sequential administration.
- the eleventh aspect of the present invention provides a use of the cell preparation according to the second aspect of the present invention, the kit according to the third aspect of the present invention, or the composition according to the seventh aspect of the present invention, for the preparation of Drugs to prevent and/or treat inflammatory diseases.
- FIG. 1 shows that low-dose IGF2 inhibits peritonitis.
- Low-dose IGF2 (L-IGF2, 5-50ng IGF2 per mouse) and high-dose IGF2 (H-IGF2, 1000ng IGF2 per mouse) were used to treat peritonitis mice.
- FIG. 2 shows that low-dose IGF2 changed the energy metabolism preference of macrophages and the expression trend and potential of inflammatory factors.
- Low-dose IGF2 L-IGF2, 5-50ng IGF2 per mouse
- high-dose IGF2 H-IGF2, 1000ng IGF2 per mouse
- the peritonitis mouse peritoneal macrophages were isolated and analyzed.
- A-B low-dose IGF2 enhances the oxidative phosphorylation potential of macrophages, while high-dose IGF2 inhibits oxidative phosphorylation.
- C low-dose IGF2 changed the energy metabolism preference of macrophages and the expression trend and potential of inflammatory factors.
- low-dose IGF2 reduces the amount of lactic acid produced by macrophages, corresponding to weaker aerobic glycolytic metabolism.
- D low-dose IGF2 inhibits the expression of LPS-induced pro-inflammatory genes (Cxcl10, IL-18, Tnfa, Cd40),
- E low-dose IGF2 promotes IL-4 and IL-13-induced anti-inflammatory repair related genes (Retnla, Ym1, Ccl22, Clec7a) expression.
- the experiment was repeated three times, and the significant difference was completed by student's t test. *P ⁇ 0.05; **P ⁇ 0.01; ***P ⁇ 0.001.
- FIG. 3 shows that low-dose IGF2 increases the cytoplasmic H + concentration of monocytes (maturing) and macrophages (mature) and reduces the lysosomal H + concentration.
- Low-dose IGF2 L-IGF2, 5-50ng IGF2 per mouse
- high-dose IGF2 H-IGF2, 1000ng IGF2 per mouse
- IGF2 Abdominal monocytes and macrophages of peritonitis mice were separated and analyzed.
- A Flow cytometric detection of IGF2 regulates the fluorescence intensity of peritoneal monocytes and macrophages JC1, and evaluates the relative mitochondrial membrane potential.
- B Flow cytometry IGF2 regulates the cytoplasmic pH of peritoneal monocytes and macrophages.
- C Flow cytometry IGF2 regulates the relative pH value of lysosomes of peritoneal macrophages.
- D IGF2 regulates the absolute activity of lysosomal type V ATPase of peritoneal macrophages.
- Figure 4 shows the construction of igf2r gene-deficient mice and igf1r or igf2r conditional knockout mice.
- AC using CRISPR-Cas9 technology to knock down IGF2R gene expression (complete knockout of IGF2R gene, lethal mouse embryos) by causing frameshift mutations caused by 2bp or 13bp gene deletion, using gene sequencing technology to identify offspring mice, using Flow cytometry to identify knockdown efficiency.
- DI by crossing IGF1R fl/fl or IGF2R fl/fl mice and Lyz Cre mice to obtain mice that specifically knock out myeloid cells IGF1R or IGF2R. The knockout efficiency was identified based on PCR and flow analysis.
- lane 1 indicates wild-type mice (124bp)
- lane 2 indicates Igf1r fl/fl (220bp)
- lane 3 has no band, indicating that Igf1r fl/fl mice do not have Lyz2 Cre activity
- Lane 4 (320bp) indicates that Igf1r fl/fl mice possess Lyz2 Cre activity.
- H Lane 1 indicates wild-type mice (2000 bp)
- Lane 2 indicates Igf2r fl/fl mice without Lyz2 Cre activity (2200 bp).
- FIG. 5 shows that low-dose IGF2 inhibits dextran sulfate sodium salt (DSS)-induced inflammatory bowel disease.
- DSS dextran sulfate sodium salt
- a mouse model of enteritis was induced by feeding and drinking 4% DSS solution.
- A-E low-dose IGF2 alleviates DSS-induced inflammatory bowel disease, which is manifested as a significant improvement in body mass index, survival time, stool score and stool bleeding score, and colon length.
- F-H and IGF2 were used to treat DSS-induced colitis mice, H&E staining, TUNEL and Ki67 immunohistochemical staining of mouse colon tissue. The experiment was repeated three times, and the significant difference was completed by student's t test.
- Figure 6 shows that macrophages treated with low-dose IGF2 can effectively inhibit inflammatory bowel disease.
- mice drinking 4% DSS solution to induce a mouse model of inflammatory bowel disease.
- A-B low-dose or high-dose IGF2-induced peritoneal mononuclear/macrophages (peritoneal macrophages, PM) in the treatment of DSS-induced inflammatory bowel disease, body weight changes and survival curves were recorded. The experiment was repeated three times, and the significant difference was completed by student's t test. **P ⁇ 0.01.
- FIG. 7 shows that low-dose IGF2 promotes macrophage formation with anti-inflammatory properties.
- A Low-dose IGF2 improves DSS-induced inflammatory bowel disease and reduces the infiltration of mononuclear cells in colon tissue.
- BC analysis of the effect of IGF2 on the expression of interleukin-1 ⁇ (IL-1 ⁇ ) by macrophages (CD11b + F4/80 +) infiltrating the colon of mice with inflammatory bowel disease.
- D Analysis of the effect of IGF2 on the expression of PD-L1 on macrophages (CD11b + F4/80 +) infiltrating the colon of mice with inflammatory bowel disease. The experiment was repeated three times, and the significant difference was completed by the student's t test. ***P ⁇ 0.001.
- FIG. 8 shows that IGF1 aggravates the progression of inflammatory bowel disease.
- mice drinking 4% DSS solution to induce a mouse model of inflammatory bowel disease.
- mice drinking 4% DSS solution to induce a mouse model of inflammatory bowel disease.
- mice with inflammatory bowel disease were injected with low-dose IGF1 (L-IGF1, 50ng/mouse) and high-dose IGF1 (H-IGF1, 1000ng/mouse) Mouse) or PBS.
- A observe the survival of mice
- B measure the changes in colon length
- C use flow cytometry to detect the proportion of IL-1 ⁇ expressed by macrophages in the colon lamina propria
- D use flow cytometry to detect the colon lamina
- Macrophages express the level of PD-L1
- EG peritonitis mice were injected with L-IGF1, H-IGF1 or PBS, and macrophages were separated from the peritoneal lavage fluid. Under LPS stimulation, IGF1 was analyzed for macrophages The effects of nitric oxide secretion, lactic acid production and glucose consumption.
- FIG. 9 shows the effect of low-dose IGF2 performed by IGF2R on the anti-inflammatory function of macrophages.
- Wild-type (WT) mice and IGF2RCKO (IGF2R fl/fl Lyz2 Cre ) mice were given to induce peritonitis, IGF2 (50ng/mouse) or PBS was intraperitoneally injected, and macrophages were separated from the peritoneal lavage fluid 48 hours later.
- A Flow cytometric analysis of the expression of PD-L1 on macrophages;
- B Flow cytometric analysis of the expression of IL-1 ⁇ in macrophages. Mice were given to induce peritonitis, low-dose IGF2 or high-dose IGF2 were injected respectively, and IGF2R antibody was given at the same time,
- C flow cytometry analysis of H + concentration in macrophage cytoplasm.
- FIG. 10 shows that IGF2R performs the anti-inflammatory task of low-dose IGF2.
- IGF2RCKO IGF2R fl/fl Lyz2 Cre
- IGF1RCKO IGF1R fl/fl Lyz2 Cre
- A weight change of mice
- B survival of mice
- C stool score
- D stool blood score
- E colonic histopathological analysis
- F mononuclear cell infiltration in the lamina intestinal of the colon.
- GL evaluation of high-dose IGF2 on the inflammatory bowel induced in IGF1R fl/fl Lyz2 Cre mice and IGF1R fl/fl mice Influence of disease.
- L mononuclear cell infiltration in the lamina limbal of the colon.
- FIG 11 shows that Leu27-IGF2 targeted activation of IGF2R optimally inhibits peritonitis and inflammatory bowel disease.
- Peritonitis mice were treated with low-dose IGF2 or Leu27-IGF2 (50ng IGF2/Leu27-IGF2 per mouse) and high-dose IGF2 or Leu27-IGF2 (1000ng, or 2000ng IGF2/Leu27-IGF2 per mouse), respectively.
- Abdominal monocytes and macrophages of peritonitis mice were separated and analyzed.
- Leu27-IGF2 significantly increases the H + concentration in the cytoplasm of macrophages.
- HJ stool score, degree of blood in the stool, and colon length evaluated the therapeutic effect of Leu27-IGF2 on DSS-induced inflammatory bowel disease.
- K Schematic diagram of the effective threshold of wild-type IGF2 and Leu27-IGF2 to relieve Ftg-induced peritonitis and DSS-induced inflammatory bowel disease. The experiment was repeated three times, and the significant difference was completed by the student's t test. *P ⁇ 0.05;**P ⁇ 0.01;***P ⁇ 0.001.
- Figure 12 shows that Leu27-Des(62-67)-IGF2 inhibits inflammatory bowel disease.
- Mice were given 4% DSS solution to induce an inflammatory bowel disease model, and low and high doses of Leu27-Des(62-67)-IGF2 were injected on the first day of disease induction.
- A monitoring the weight change of mice;
- B survival of mice with inflammatory bowel disease, evaluating the therapeutic effect of Leu27-Des(62-67) IGF2 on inflammatory bowel disease.
- Significant differences are completed by student’s t test. *P ⁇ 0.05; ***P ⁇ 0.001.
- FIG 13 shows that activation of IGF2R promotes nuclear distribution and regulates the cytoplasmic pH of macrophages.
- THP1 cells were treated with different doses of IGF2, AB, and flow cytometry was used to detect the presence of IGF2R on the cell membrane surface.
- C Western blotting detects the expression of IGF2R at the overall cell level and in the nucleus.
- the protein nuclear transcription inhibitor-ivermectin (5 ⁇ M) was used to treat bone marrow-derived mononuclear precursor cells treated with different doses of IGF2. After the macrophages matured, D, Western blotting was used to detect the expression of IGF2R at the overall cell level and in the nucleus.
- THP1 cells were treated with different doses of IGF2 or Leu27-Des(62-67)-IGF2, F, flow cytometry analysis technique was used to detect the expression of IGF2R on the cell membrane surface. Significant differences are completed by student's t test. ***P ⁇ 0.001.
- non-IGF1R-binding substances can significantly (a) prevent and/or treat inflammatory diseases; and/or (b) prevent and/or treat autoimmune diseases .
- the inventors completed the present invention.
- wild-type IGF2 wild-type IGF2 amino acid fragments from positions 25 to 91, glutamic acid at position 12 is mutated to Asp, Ala, Gln, His, Arg, or Lys, IGF2 mutant, phenylalanine at position 26
- the IGF2 mutant with Ser, the IGF2 mutant with tyrosine 27 mutated to Leu, the IGF2 mutant with valine 43 mutated to Leu, and the IGF2 mutant with D-domain deleted are in SEQ ID NO.:1
- the truncation, mutation and/or deletion of amino acids are carried out on the basis of.
- IGF2 protein and its mutants
- IGF2 Insulin like growth factor 2
- IGF2 receptors include IGF1R and IGF2R, among which IGF2R has the highest affinity, followed by binding to IGF1R, and almost no binding to insulin receptor 2 .
- IGF2 and IGF1 have a high degree of homology in amino acid sequence, with 62% overlapping sequences. Therefore, IGF2 can often reflect the biological effects that IGF1 can activate 2 .
- Current research suggests that the main biological function of IGF2 depends on the activation of tyrosine kinases that activate IGF1R (insulin like growth factor 1 receptor), but has no positive correlation with IGF2R (insulin like growth factor 2 receptor) and IR. From an evolutionary perspective, IGF1R and IR have gene homology, and the signaling pathways activated downstream of IGF1R and IR are very similar. However, the source of IGF2R gene is completely different from that of IGF1R or IR gene.
- IGF2R is often considered to have the opposite biological function of IGF1R and IR.
- IGF2R acts as the degradation receptor of IGF2, and inhibits IGF2's effect on IGF1R by internalizing degradation of IGF2. The activation effect.
- IGF2R Intracellular Golgi apparatus
- IGF2R is not the only receptor of IGF2
- IGF2 is not the only ligand of IGF2R. Its ligands also include mannose-6-phosphate.
- the main function of IGF2R is to transport its ligands to lysosomes. 4 complete protein processing or degradation.
- the IGF2R on the cell membrane can be cut off in a soluble form, into the blood and other liquid environment, effective binding IGF2, 4,5 inhibition IGF2 function.
- IGF2 can inhibit animal models of multiple sclerosis and ulcerative colitis, but its specific regulation mechanism is still unclear.
- the present invention relates to an IGF2 protein and variants thereof.
- the amino acid sequence of the IGF2 protein is shown in SEQ ID NO.:1.
- the IGF2 protein or the vector expressing the IGF2 protein of the present invention can (a) prevent and/or treat inflammatory diseases.
- the IGF2 protein or the vector expressing the IGF2 protein of the present invention can also be used for one or more purposes selected from the following group:
- the present invention also includes 50% or more of the sequence shown in SEQ ID NO. 1 of the present invention (preferably 60% or more, 70% or more, 80% or more, more preferably 90% or more, more preferably 95% or more, most preferably 98% or more, such as 99%) homologous polypeptides or proteins with the same or similar functions.
- SEQ ID NO.: 1 is a human IGF2 protein (AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRGIVEECCFRSCDLALLETYCATPAKSE).
- the "same or similar function” mainly refers to: "(a) prevention and/or treatment of inflammatory diseases; and/or (i) reduction of pathogenic mononuclear cell infiltration; and/or (ii) inhibition of macrophage Pathogenic phenotype; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the cytoplasmic H + concentration of macrophages; and/or (v) increase the metabolic propensity of macrophages to oxidize phosphoric acid ; And/or (vi) endowing macrophages with immune memory that significantly inhibits inflammation; and/or (vii) preventing and/or treating autoimmune diseases; and/or (viii) a variety of other macrophages involved Diseases involving inflammation (such as inflammation related to liver fibrosis, renal fibrosis, lung fibrosis and other diseases, inflammation caused by tissue, organ or cell transplantation, and lung cancer, breast cancer, prostate cancer, liver cancer , Pancreatic cancer,
- the protein of the present invention can be a recombinant protein, a natural protein, or a synthetic protein.
- the protein of the present invention can be a natural purified product, or a chemically synthesized product, or produced from a prokaryotic or eukaryotic host (for example, bacteria, yeast, higher plant, insect, and mammalian cells) using recombinant technology.
- a prokaryotic or eukaryotic host for example, bacteria, yeast, higher plant, insect, and mammalian cells
- the protein of the present invention may be glycosylated or non-glycosylated.
- the protein of the present invention may also include or not include the initial methionine residue.
- the present invention also includes IGF2 protein fragments and analogs having IGF2 protein activity.
- fragment and analogs having IGF2 protein activity refer to a protein that substantially maintains the same biological function or activity as the natural IGF2 protein of the present invention.
- the mutant of the IGF2 protein of the present invention is mutated in the tyrosine at position 27 of the wild-type IGF2 protein corresponding to SEQ ID NO.:1 (AYRPSETLCGGELVDTLQFVCGDRGFYFSRPASRVSRRSRGIVEECCFRSCDLALLETYCATPAKSE), and the IGF2 protein of the present invention is Mutants can significantly (a) prevent and/or treat inflammatory diseases; and/or (b) prevent and/or treat autoimmune diseases.
- the tyrosine at position 27 is mutated to leucine.
- the tyrosine at position 27 is mutated to leucine, and D-domain (threonine at position 62 to glutamic acid at position 67 of SEQ ID NO.: 1) is optionally deleted.
- the IGF2 mutant is a polypeptide having an amino acid sequence shown in any one of SEQ ID NO.: 2-58, an active fragment thereof, or a conservative variant polypeptide thereof.
- amino acid numbering in the mutant of the present invention is based on SEQ ID NO.:1.
- amino acid of the mutant may be misaligned with respect to the amino acid numbering of SEQ ID NO.: 1), such as misaligned 1-5 to the N-terminus or C-terminus of the amino acid.
- the mutein fragment, derivative or analogue of the present invention may be (i) a mutein in which one or more conservative or non-conservative amino acid residues (preferably conservative amino acid residues) are substituted, and such substituted amino acids
- the residue may or may not be encoded by the genetic code, or (ii) a mutein with a substitution group in one or more amino acid residues, or (iii) a mature mutein and another compound (such as an extended mutein) Half-life compounds, such as polyethylene glycol) fused to form a mutant protein, or (iv) additional amino acid sequence fused to the mutant protein sequence to form a mutant protein (such as leader sequence or secretory sequence or used to purify the mutant protein)
- the sequence or proprotein sequence, or the fusion protein formed with the antigen IgG fragment According to the teachings herein, these fragments, derivatives and analogs belong to the scope well known to those skilled in the art.
- conservatively substituted amino acids are preferably generated by amino acid substitution
- the present invention also includes that the natural IGF2 protein of the present invention has 50% or more (preferably 60% or more, 70% or more, 80% or more, more preferably 90% or more, more preferably 95% or more, most preferably 98% or more, such as 99 %) Homologous polypeptides or proteins with the same or similar functions.
- the protein variant can be obtained by substituting, deleting or adding at least one amino acid by several (usually 1-60, preferably 1-30, more preferably 1-20, most preferably 1-10). Derivative sequences, and adding one or several (usually within 20, preferably within 10, more preferably within 5) amino acids at the C-terminal and/or N-terminal.
- the function of the protein is usually not changed, and the addition of one or several amino acids to the C-terminal and/or ⁇ terminal usually does not change the function of the protein.
- the present invention includes that the difference between the natural IGF2 protein analog and the natural IGF2 protein may be the difference in the amino acid sequence, the difference in the modified form that does not affect the sequence, or both.
- Analogs of these proteins include natural or induced genetic variants. Induced variants can be obtained by various techniques, such as random mutagenesis by radiation or exposure to mutagens, site-directed mutagenesis or other known biological techniques.
- Analogs also include analogs having residues different from natural L-amino acids (such as D-amino acids), and analogs having non-naturally occurring or synthetic amino acids (such as ⁇ , ⁇ -amino acids). It should be understood that the protein of the present invention is not limited to the representative proteins exemplified above.
- Modified (usually not changing the primary structure) forms include: chemically derived forms of proteins in vivo or in vitro, such as acetylation or carboxylation. Modifications also include glycosylation, such as those that undergo glycosylation modifications during protein synthesis and processing. This modification can be accomplished by exposing the protein to an enzyme that performs glycosylation (such as a mammalian glycosylase or deglycosylase). Modified forms also include sequences with phosphorylated amino acid residues (such as phosphotyrosine, phosphoserine, and phosphothreonine). In addition, the mutant protein of the present invention can also be modified.
- Modified (usually not changing the primary structure) forms include: chemically derived forms of mutein in vivo or in vitro, such as acetylation or carboxylation. Modifications also include glycosylation, such as those produced by glycosylation modifications during the synthesis and processing of the mutant protein or during further processing steps. This modification can be accomplished by exposing the mutein to an enzyme that performs glycosylation, such as a mammalian glycosylase or deglycosylase. Modified forms also include sequences with phosphorylated amino acid residues (such as phosphotyrosine, phosphoserine, and phosphothreonine). It also includes mutant proteins that have been modified to improve their resistance to proteolysis or optimize their solubility.
- the present invention also provides a polynucleotide sequence encoding IGF2 protein.
- the polynucleotide of the present invention may be in the form of DNA or RNA.
- DNA forms include: DNA, genomic DNA or synthetic DNA.
- DNA can be single-stranded or double-stranded.
- a polynucleotide encoding a mature polypeptide includes: a coding sequence that only encodes the mature polypeptide; the coding sequence of the mature polypeptide and various additional coding sequences; the coding sequence (and optional additional coding sequences) of the mature polypeptide and non-coding sequences.
- polynucleotide encoding a polypeptide may include a polynucleotide encoding the polypeptide, or a polynucleotide that also includes additional coding and/or non-coding sequences.
- the present invention also relates to variants of the aforementioned polynucleotides, which encode fragments, analogs and derivatives of polypeptides having the same amino acid sequence as the present invention.
- the variants of this polynucleotide can be naturally occurring allelic variants or non-naturally occurring variants.
- These nucleotide variants include substitution variants, deletion variants and insertion variants.
- an allelic variant is an alternative form of a polynucleotide. It may be a substitution, deletion or insertion of one or more nucleotides, but it will not substantially change the function of the encoded polypeptide. .
- the coding nucleic acid sequence of the present invention can be constructed by the method of synthesizing the nucleotide sequence in segments and then performing overlap extension PCR.
- the present invention also relates to polynucleotides that hybridize with the aforementioned sequences and have at least 50%, preferably at least 70%, and more preferably at least 80% identity between the two sequences.
- the present invention particularly relates to polynucleotides that can hybridize with the polynucleotide of the present invention under stringent conditions (or stringent conditions).
- stringent conditions refer to: (1) hybridization and elution at lower ionic strength and higher temperature, such as 0.2 ⁇ SSC, 0.1% SDS, 60°C; or (2) adding during hybridization There are denaturants, such as 50% (v/v) formamide, 0.1% calf serum/0.1% Ficoll, 42°C, etc.; or (3) only the identity between the two sequences is at least 90% or more, more Fortunately, hybridization occurs when more than 95%.
- proteins and polynucleotides of the present invention are preferably provided in an isolated form, and more preferably, are purified to homogeneity.
- the full-length sequence of the polynucleotide of the present invention can usually be obtained by PCR amplification method, recombination method or artificial synthesis method.
- primers can be designed according to the relevant nucleotide sequence disclosed in the present invention, especially the open reading frame sequence, and a commercially available cDNA library or a cDNA prepared by a conventional method known to those skilled in the art can be used.
- the library is used as a template to amplify the relevant sequences. When the sequence is long, it is often necessary to perform two or more PCR amplifications, and then splice the amplified fragments together in the correct order.
- the recombination method can be used to obtain the relevant sequence in large quantities. This is usually done by cloning it into a vector, then transferring it into a cell, and then isolating the relevant sequence from the proliferated host cell by conventional methods.
- artificial synthesis methods can also be used to synthesize related sequences, especially when the fragment length is short. Usually, by first synthesizing multiple small fragments, and then ligating to obtain fragments with very long sequences.
- the DNA sequence encoding the protein (or fragment or derivative thereof) of the present invention can be obtained completely through chemical synthesis.
- the DNA sequence can then be introduced into various existing DNA molecules (or such as vectors) and cells known in the art.
- mutations can also be introduced into the protein sequence of the present invention through chemical synthesis.
- the method of using PCR technology to amplify DNA/RNA is preferably used to obtain the polynucleotide of the present invention.
- the RACE method RACE-cDNA end rapid amplification method
- the primers used for PCR can be appropriately selected according to the sequence information of the present invention disclosed herein. And can be synthesized by conventional methods.
- the amplified DNA/RNA fragments can be separated and purified by conventional methods such as gel electrophoresis.
- the present invention also relates to a vector containing the polynucleotide of the present invention, a host cell produced by genetic engineering using the vector of the present invention or a mutant protein coding sequence of the present invention, and a method for producing the polypeptide of the present invention through recombinant technology.
- the polynucleotide sequence of the present invention can be used to express or produce recombinant mutein. Generally speaking, there are the following steps:
- polynucleotide (or variant) of the present invention encoding the mutant protein of the present invention, or use a recombinant expression vector containing the polynucleotide to transform or transduce a suitable host cell;
- the polynucleotide sequence encoding the mutant protein can be inserted into a recombinant expression vector.
- recombinant expression vector refers to bacterial plasmids, bacteriophages, yeast plasmids, plant cell viruses, mammalian cell viruses such as adenovirus, retrovirus or other vectors well known in the art. Any plasmid and vector can be used as long as it can replicate and stabilize in the host.
- An important feature of an expression vector is that it usually contains an origin of replication, a promoter, a marker gene, and translation control elements.
- Methods well known to those skilled in the art can be used to construct an expression vector containing the DNA sequence encoding the mutein of the present invention and appropriate transcription/translation control signals. These methods include in vitro recombinant DNA technology, DNA synthesis technology, and in vivo recombination technology.
- the DNA sequence can be effectively linked to an appropriate promoter in the expression vector to guide mRNA synthesis.
- promoters are: Escherichia coli lac or trp promoter; lambda phage PL promoter; eukaryotic promoters include CMV immediate early promoter, HSV thymidine kinase promoter, early and late SV40 promoter, anti Transcriptional virus LTRs and some other known promoters that can control gene expression in prokaryotic or eukaryotic cells or viruses.
- the expression vector also includes a ribosome binding site for translation initiation and a transcription terminator.
- the expression vector preferably contains one or more selectable marker genes to provide phenotypic traits for selecting transformed host cells, such as dihydrofolate reductase for eukaryotic cell culture, neomycin resistance, and green Fluorescent protein (GFP), or tetracycline or ampicillin resistance for E. coli.
- selectable marker genes to provide phenotypic traits for selecting transformed host cells, such as dihydrofolate reductase for eukaryotic cell culture, neomycin resistance, and green Fluorescent protein (GFP), or tetracycline or ampicillin resistance for E. coli.
- a vector containing the above-mentioned appropriate DNA sequence and an appropriate promoter or control sequence can be used to transform an appropriate host cell so that it can express the protein.
- the host cell can be a prokaryotic cell, such as a bacterial cell; or a lower eukaryotic cell, such as a yeast cell; or a higher eukaryotic cell, such as a mammalian cell.
- a prokaryotic cell such as a bacterial cell
- a lower eukaryotic cell such as a yeast cell
- a higher eukaryotic cell such as a mammalian cell.
- Representative examples include: Escherichia coli, Streptomyces; bacterial cells of Salmonella typhimurium; fungal cells such as yeast and plant cells (such as ginseng cells).
- Enhancers are cis-acting factors of DNA, usually about 10 to 300 base pairs, acting on promoters to enhance gene transcription. Examples include the 100 to 270 base pair SV40 enhancer on the late side of the replication initiation point, the polyoma enhancer on the late side of the replication initiation point, and adenovirus enhancers.
- Transformation of host cells with recombinant DNA can be performed by conventional techniques well known to those skilled in the art.
- the host is a prokaryotic organism such as Escherichia coli
- competent cells that can absorb DNA can be harvested after the exponential growth phase and treated with the CaCl2 method.
- the steps used are well known in the art.
- Another method is to use MgCl2.
- transformation can also be carried out by electroporation.
- the following DNA transfection methods can be selected: calcium phosphate co-precipitation method, conventional mechanical methods such as microinjection, electroporation, liposome packaging, etc.
- the obtained transformants can be cultured by conventional methods to express the polypeptide encoded by the gene of the present invention.
- the medium used in the culture can be selected from various conventional mediums.
- the culture is carried out under conditions suitable for the growth of the host cell. After the host cells have grown to an appropriate cell density, the selected promoter is induced by a suitable method (such as temperature conversion or chemical induction), and the cells are cultured for a period of time.
- the recombinant polypeptide in the above method can be expressed in the cell or on the cell membrane, or secreted out of the cell. If necessary, the physical, chemical, and other characteristics can be used to separate and purify the recombinant protein through various separation methods. These methods are well known to those skilled in the art. Examples of these methods include, but are not limited to: conventional renaturation treatment, treatment with a protein precipitation agent (salting out method), centrifugation, osmotic sterilization, ultra-treatment, ultra-centrifugation, molecular sieve chromatography (gel filtration), adsorption layer Analysis, ion exchange chromatography, high performance liquid chromatography (HPLC) and various other liquid chromatography techniques and combinations of these methods.
- non-IGF1R binding type As used herein, the terms “non-IGF1R binding type”, “IGF2R selective type” or “IGF2R selective” are used interchangeably, and refer to the substance that is more selective for IGF2R than for IGF1R, that is, selectively and/ Or preferentially bind to IGF2R, but not or substantially not bind to IGF1R.
- non-IGF1R-binding substances include substances that have a higher selectivity or affinity for IGF2R than IGF1R, and substances that can efficiently activate IGF2R and do not activate IGF1R (such as non-IGF1R ligands such as retinoic acid).
- the affinity or selectivity of the non-IGF1R-binding substance for IGF2R is higher than its affinity or selectivity for IGF1R.
- the non-IGF1R-binding substance is selected from the group consisting of IGF2 mutants, vectors expressing IGF2 mutants, antibodies, small molecule compounds, or combinations thereof.
- the non-IGF1R-binding substance also includes one or more substances selected from the following group: Plasminogen, Serglycin, e1a stimulator Cellular suppressor of E1A-stimulated genes (CREG), Sulfamidase (Sgsh), phosphorylated ⁇ -glucuronidase (Phosphorylated ⁇ -glucuronidase, Gusb) and other effective anti-inflammatory ligands for IGF2R .
- Plasminogen Plasminogen, Serglycin, e1a stimulator Cellular suppressor of E1A-stimulated genes (CREG), Sulfamidase (Sgsh), phosphorylated ⁇ -glucuronidase (Phosphorylated ⁇ -glucuronidase, Gusb) and other effective anti-inflammatory ligands for IGF2R .
- non-IGF1R-binding substances can significantly (a) prevent and/or treat inflammatory diseases; and/or (i) reduce disease-causing mononuclear cell infiltration; and/or (ii) Inhibit the pathogenic phenotype of macrophages; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the cytoplasmic H + concentration of macrophages; and/or (v) increase macrophage oxidation
- the metabolic propensity of phosphoric acid; and/or (vi) confers immune memory to macrophages that significantly inhibits inflammation; and/or (vii) prevents and/or treats autoimmune diseases; and/or (viii) is used for macrophages Participate in a variety of other diseases involving inflammation (such as inflammation related to liver fibrosis, renal fibrosis, lung fibrosis and other diseases, inflammation related to tissue, organ or cell transplantation, and lung cancer, breast cancer
- the present invention provides a compound pharmaceutical composition containing active ingredients (a) phagocytic cells treated with a non-IGF1R-binding substance; (b) optional non-IGF1R-binding substance; and (c) a pharmaceutically acceptable carrier .
- Such carriers include (but are not limited to): saline, buffer, dextrose, water, glycerol, ethanol, powder, and combinations thereof.
- the pharmaceutical preparation should match the mode of administration.
- the pharmaceutical composition of the present invention can be made into an injection form, for example, prepared by conventional methods with physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods.
- compositions such as injections, solutions, tablets and capsules should be manufactured under sterile conditions.
- the pharmaceutical combination of the present invention can also be made into powder for inhalation.
- the dosage form of the pharmaceutical composition of the present invention is an injection.
- the amount of active ingredient administered is a therapeutically effective amount.
- the pharmaceutical preparation of the present invention can also be made into a sustained-release preparation.
- the pharmaceutical composition of the present invention is preferably an injection preparation.
- the pharmaceutical composition of the present invention can also be used together with other therapeutic agents.
- the pharmaceutical composition of the present invention may also include additional components selected from the following group: (a) drugs for preventing and/or treating inflammatory diseases; and/or (b) preventing and/ Or a component for the treatment of autoimmune diseases.
- the effective amount of the active ingredient of the present invention can vary with the mode of administration and the severity of the disease to be treated.
- the selection of the preferred effective amount can be determined by a person of ordinary skill in the art according to various factors (for example, through clinical trials).
- the factors include, but are not limited to: the pharmacokinetic parameters of the active ingredients such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated by the patient, the patient's weight, the patient's immune status, and administration The way and so on.
- the active ingredient of the present invention is about 0.01 ⁇ g-1 mg/kg body weight per day, preferably 0.1-10 ⁇ g/kg body weight, more preferably 0.5-5 ⁇ g/kg body weight. It can be given satisfactory results.
- the optimal dose will be adjusted according to the course and condition of the disease. For example, according to the urgent requirement of the treatment condition, the dose can be given several times a day, or the dose can be reduced proportionally.
- the pharmaceutically acceptable carriers of the present invention include (but are not limited to): water, saline, liposomes, lipids, proteins, protein-antibody conjugates, peptides, cellulose, nanogels, or Its combination.
- the choice of carrier should match the mode of administration, which are well known to those of ordinary skill in the art.
- the present invention also provides a drug that can be used to (a) prevent and/or treat inflammatory diseases; and/or ((i) reduce the infiltration of pathogenic mononuclear cells; and/or (ii) inhibit the infiltration of macrophages Disease phenotype; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the cytoplasmic H + concentration of macrophages; and/or (v) increase the metabolic propensity of macrophages to oxidize phosphoric acid; (vi) Endow macrophages with immune memory that significantly inhibits inflammation; and/or (vii) prevent and/or treat autoimmune diseases; and/or (viii) use for a variety of other inflammatory reactions involving macrophages Diseases (such as inflammation related to liver fibrosis, renal fibrosis, lung fibrosis and other diseases, inflammation related to tissue, organ or cell transplantation, and lung cancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer, Inflammation
- the pharmaceutical composition and kit of the present invention are suitable for (a) preventing and/or treating inflammatory diseases; and/or (i) reducing the infiltration of pathogenic mononuclear cells in the colon; and/or (ii) inhibiting macrophage Pathogenic phenotype; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the cytoplasmic H + concentration of macrophages; and/or (v) increase the metabolic propensity of macrophages to oxidize phosphoric acid ; And/or (vi) endowing macrophages with immune memory that significantly inhibits inflammation; and/or (vii) preventing and/or treating autoimmune diseases; and/or (viii) a variety of other macrophages involved Diseases involving inflammation (such as inflammation related to liver fibrosis, renal fibrosis, lung fibrosis and other diseases, inflammation caused by tissue, organ or cell transplantation, and lung cancer, breast cancer, prostate cancer, liver cancer , Pancreatic cancer,
- the preparation of the present invention can be taken three times a day to once every ten days, or once every ten days in a sustained-release manner.
- the preferred way is to take it once a day, because it facilitates the patient's adherence, thereby significantly improving the patient's compliance with medication.
- the total dose applied daily in most cases should be lower (or equal to or slightly greater than) the daily daily dose of each single drug in most cases.
- the effective dose of the active ingredient used can vary depending on the mode of administration and the expected dose. The severity of the disease to be treated varies.
- the present invention also provides the use of the two active ingredients of the present invention or corresponding drugs to (a) prevent and/or treat inflammatory diseases; and/or (i) reduce the infiltration of pathogenic mononuclear cells in the colon; and/or ( ii) inhibit the pathogenic phenotype of macrophages; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the H + concentration of the cytoplasm of macrophages; and/or (v) increase macrophages
- the two active ingredients of the present invention can be mixed with one or more pharmaceutically acceptable carriers or excipients, such as solvents, diluents, etc., and can be administered orally in the form of tablets: Pills, pills, capsules, dispersible powders, granules or suspensions (containing, for example, about 0.05-5% suspending agent), syrups (containing, for example, about 10-50% sugar), and elixirs (containing about 20-50% ethanol), Or in the form of a sterile injectable solution or suspension (containing about 0.05-5% suspension in an isotonic medium) for parenteral administration.
- these pharmaceutical preparations may contain about 0.01-99%, more preferably about 0.1%-90% by weight of the active ingredient mixed with the carrier.
- the two active ingredients or pharmaceutical compositions of the present invention can be administered by conventional routes, including (but not limited to): intramuscular, intraperitoneal, intravenous, subcutaneous, intradermal, oral, intratumoral or topical administration .
- routes of administration include oral administration, intramuscular administration or intravenous administration.
- the preferred pharmaceutical composition is a liquid composition, especially an injection.
- the two active ingredients or drugs of the present invention can also be combined with other (a) prevention and/or treatment of inflammatory diseases; and/or (i) reduction of pathogenic mononuclear cell infiltration; and/or (ii) inhibition of macrophages
- non-IGF1R-binding substances can significantly (a) prevent and/or treat inflammatory diseases; and/or (i) reduce the infiltration of pathogenic mononuclear cells; and/or (ii) inhibit macrophage The pathogenic phenotype of phages; and/or (iii) reduce the number of macrophages infiltrated; and/or (iv) increase the cytoplasmic H + concentration of macrophages; and/or (v) increase the oxidative phosphoric acid of macrophages Metabolic propensity; and/or (vi) endowed macrophages with immune memory that significantly inhibits inflammation; and/or (vii) prevention and/or treatment of autoimmune diseases; and/or (viii) used for macrophage participation
- inflammation such as inflammation related to liver fibrosis, renal fibrosis, lung fibrosis and other diseases, inflammation related to tissue, organ or cell transplantation, and lung cancer, breast cancer, prostate Inflammation
- IGF2 insulin-like growth factor 2
- IGF2R IGF2 receptor
- the present invention finds for the first time that low-dose IGF2 can induce non-classical mitochondrial dynamics and give macrophages immune memory against inflammation.
- High-dose IGF2 can activate IGF1 receptor (IGF1R), inhibit the effect of IGF2R, and promote monotherapy.
- IGF1R IGF1 receptor
- Nuclear cells and macrophages use aerobic glycolysis and metabolism to obtain pro-inflammatory capabilities.
- the present invention finds for the first time that IGF2 mutants with low affinity to IGF1R specifically activate IGF2R, and to the greatest extent alleviate autoimmune diseases and inflammatory diseases such as peritonitis and inflammatory bowel disease. These findings indicate that the targeted activation of IGF2R can determine the long-term immunosuppressive memory of macrophages. IGF2R is an ideal target for the treatment of inflammation-related diseases involving macrophages.
- the present invention discovered for the first time that the IGF2R, which is directed to activate monocytes, inhibits inflammatory bowel disease.
- the present invention found for the first time that IGF2 treatment of inflammatory bowel disease relies on macrophages.
- the present invention discovered for the first time that IGF2 is involved in a variety of other inflammatory diseases in macrophages.
- Recombinant human insulin-like growth factor-2 (Recombinant human IGF2, 292-G2-250), recombinant mouse-derived macrophage colony stimulating factor (Recombinant mouse M-CSF protein, 416-ML-050) were purchased from R&D Systems.
- the 27th human source is leucine point mutation IGF2 (Human Leu27-IGF2, TU100) purchased from GroPep Bioreagents.
- the 27th human source is leucine point mutation IGF2 (Human Leu27-IGF2, TU100). The deletion of the D domain is synthesized in the laboratory.
- IGF2R +/- mice use the Crisper-Cas9 technology to knock out the 2bp or 13bp base on the 6th exon of the igf2r gene, causing a frameshift mutation. Since homozygous IGF2R deficiency can cause embryonic death, we use IGF2R +/- heterozygous mice to pass down and identify them.
- the primers used for identification are:
- Igf2r fl/fl mice insert loxp sequence on both sides of exon 2 of igf2r gene, and the primers used for identification are:
- Primer 2,5’-CAATTTAGGGCTGAAGCGATGGAT-3’ (SEQ ID NO.: 62)
- the primers used for Igf1r fl/fl mouse identification are:
- the primers used for Lyz2 Cre mouse identification are:
- thioglycolate (Ftg) medium aqueous solution Prepare 5% (mass/volume) thioglycolate (Ftg) medium aqueous solution, sterilize at 121°C under high temperature and high pressure for 30 minutes, and cool to room temperature for later use.
- Ftg thioglycolate
- the viscosity should not cause bubbles when shaking.
- the color of the medium should be changed from blue-green when initially configured to brown when shaking.
- mice C57BL/6 female mice of the same age in the range of 9-12 weeks were selected for the experiment. Each mouse was slowly injected with 2ml of thioglycolate medium into the abdominal cavity, 24 hours later, IGF2 or other drugs were given, and it was given once at the 24th and 48th hours after the thioglycolate injection, and at the 72nd hour The mice were euthanized and the peritoneal macrophages were isolated.
- mice that have been euthanized (Ftg) for 72 hours with 75% ethanol for a few minutes, tear off the abdominal fur in the direction perpendicular to the head and tail of the mouse, suck 10ml of pre-cooled phosphate buffer () with a syringe, and remove the phosphoric acid.
- salt buffer solution into the abdominal cavity of the mouse (to help blow down the adherent cells), clamp the tail of the mouse with the index finger and ring finger of the right hand, gently hold the mouse, shake it up and down three to five times, and slowly use a syringe Aspirate the cell lavage fluid.
- Peritoneal macrophage lavage fluid needs to be stored at four degrees, and cells are easy to adsorb to the wall of the centrifuge tube at room temperature.
- DSS dextran sulfate sodium salt
- the clinical scoring of inflammatory bowel disease was performed according to the following criteria. After feeding the mice with 4% DSS solution for 4 days, “Stool scores” and “Bleeding scores” were evaluated.
- the "Stool scores” standard is: 0 points, normal stool shape; 1 point, semi-formed stool and no anus attachment; 2 points, semi-formed stool and anal attachment; 3 points, liquid feces attached to the anus.
- "Bleeding scores” judgment criteria 0 points, fecal occult blood test paper is negative; 1 point, fecal occult blood test paper is positive; 2 points, blood stains can be seen in the cage; 3 points, rectal bleeding, red blood spots on the anus.
- mice Euthanize the mice induced by 4% DSS for 4-5 days, dissected the mice, and obtained the colon tissue, and placed it in 4°C pre-cooled phosphate buffer solution to wash off the blood stains on the colon surface. Cut the colon longitudinally, and carefully wash away the stool in a four-degree pre-cooled phosphate buffer.
- Use 1640 culture medium containing 200 ⁇ g/ml DNaseI and 1mg/ml type VIII collagenase add DTT before digestion, shake 200 rpm for 60 minutes, carefully remove the colon basal tissue, cut into small sections, and perform digestion again , Filter with a 70 ⁇ m cell sieve, and centrifuge the filtrate at 400g for 5 minutes to obtain a single cell pellet.
- Protein staining on cell membrane surface For cells blocked with CD16/32 antibody, dilute the antibody with phosphate buffer at a ratio of 1:100, 100 ⁇ l staining working solution/1 ⁇ 10 7 cells, stain at room temperature for 30 minutes, wash with phosphate buffer, and centrifuge with 200 ⁇ l Resuspend in phosphate buffer, test on the machine or fix with 1% neutral formaldehyde.
- Intracellular proteins and nuclear proteins need to be performed after the staining of the cell membrane surface proteins is completed.
- intracellular proteins use the cell fixation and rupture kit (Fixation/Permeabilization Solution and Perm/Wash Buffer) for rupture and staining after fixation; for nuclear proteins, use the cell fixation/cell rupture kit (Foxp3/Transcription Factor) Staining Buffer Set) Stain after fixation and membrane rupture.
- the oxygen consumption rate measurement experiment and the extracellular acid production rate measurement experiment were performed with or without the following stimulation conditions: 1 ⁇ M oligomycin, 0.75mM FCCP, 100nM rotenone and 1 ⁇ M antimycin, using an extracellular flux analyzer , Agilent) determination.
- RNA Isolation Kit Use Tiangen Biological Cell/Bacterial RNA Isolation Kit to extract total RNA from cells. You can usually get 300ng/ml RNA in a volume of 30 ⁇ L per sample.
- the real-time quantitative PCR system is 10 ⁇ L, including 5 ⁇ L SYBR, 4 ⁇ L cDNA, and 1 ⁇ L primers. After spotting the sample in the 384-well plate, centrifuge it at 2500 rpm for 2 minutes at room temperature. In this step, centrifugation at 4°C will form water mist on the membrane.
- the real-time quantitative PCR conditions are: Step 1, 95°C for 10 minutes, Step 2, 95°C for 15 seconds, Step 3, 60°C for 1 minute, Step 4, 95°C for 15 seconds, Step 5, 60°C for 1 minute , Step 6, 95°C for 15 seconds, of which Step 2 and Step 3 cycle 40 times.
- the primers involved in the real-time quantitative PCR experiment are as follows:
- TNFa-F 5’-CCTGTAGCCCACGTCGTAG-3’ (SEQ ID NO.: 69),
- TNFa-R 5’-GGGAGTAGACAAGGTACAACCC-3’ (SEQ ID NO.: 70);
- IL-1b-F 5’-GCAACTGTTCCTGAACTCAACT-3’ (SEQ ID NO.: 71),
- IL-1b-R 5'-ATCTTTTGGGGTCCGTCAACT-3' (SEQ ID NO.: 72);
- Il18-F 5’-GACTCTTGCGTCAACTTCAAGG-3’ (SEQ ID NO.: 81)
- Il18-R 5’-CAGGCTGTCTTTTGTCAACGA’ (SEQ ID NO.: 82);
- Retnla-F 5’-CCAATCCAGCTAACTATCCCTCC-3’ (SEQ ID NO.: 75),
- Retnla-R 5’-ACCCAGTAGCAGTCATCCCA-3’ (SEQ ID NO.: 76);
- Dehydration embedding Take the colon tissue at the same position and fix it in 4% paraformaldehyde solution for 24 hours, and then embed it after dehydration treatment in the following steps. 70% ethanol, soak overnight; 80% ethanol, soak for 2 hours; 85% ethanol, soak for 1.5 hours; 90% ethanol, soak for 1 hour; 95% ethanol, soak for 1 hour; 100% ethanol, soak for 30 minutes, repeat the operation once ; Xylene, soak for 5 minutes, repeat the operation once; 60 °C liquid paraffin, soak for 1 hour, repeat the operation once; use the paraffin embedding machine to complete the paraffin embedding.
- Section staining slice at a thickness of 3 ⁇ m, extend the paraffin tissue section in a 56°C water bath for a few seconds, absorb the paraffin tissue section with a glass slide, bake the slice in an oven at 60°C overnight, and then perform staining according to the following steps.
- Xylene soak for 5 minutes, repeat the operation once; 100% ethanol, soak for 5 minutes, repeat the operation once; 95% ethanol, soak for 5 minutes, repeat the operation once; 70% ethanol, soak for 5 minutes; distilled water, soak for 5 minutes; Wood essence, dyeing for 5 minutes; tap water and running water for proper time; ammonia water, immersion for proper time; running water for proper time; eosin, dyeing for 1 minute; 95% ethanol, soaking for 1 minute; 100% ethanol, soaking for 1 minute, repeat the operation once.
- Xylene soak for 1 minute; repeat the operation once; mount the tablet.
- Antigen-antibody reaction drop the primary anti-Ki67 antibody on the paraffin section, incubate overnight at 4°C; wash four times with pH 7.4 phosphate-Tween buffer for three minutes each time; add horseradish on the paraffin section dropwise Peroxidase-conjugated secondary antibody, incubate at room temperature for 60 minutes; wash 4 times with pH 7.4 phosphate-Tween buffer for 3 minutes each time; after DBA staining for an appropriate time, wash off DBA stain; use hematoxylin Dye counterstain paraffin sections, determine the appropriate time according to the type of tissue; rinse with tap water to remove hematoxylin; use hydrochloric acid alcohol differentiation and ammonia water to reverse blue until hematoxylin staining appears light blue; dehydration mounts.
- the TUNEL immunohistochemistry experiment was carried out in accordance with the experimental method provided by the In situ cell death detection kit-POD kit (Roche, TUN11684817).
- Detecting the accumulation of nitric oxide in the supernatant of the culture medium can reflect the activity of inducible nitric oxide synthase (iNOS) of macrophages, and speculate the type of energy metabolism and inflammatory phenotype of macrophages.
- iNOS inducible nitric oxide synthase
- the lactate detection kit was used to detect the lactate production capacity of the peritoneal monocytes and macrophages pre-programmed by IGF2. Perform the experiment according to the experimental method provided by Actate Colorimetric Assay Kit II (BioVision, K627-100). Detecting the accumulation of lactic acid in the supernatant of the culture medium can reflect the type of energy metabolism and inflammatory phenotype of macrophages.
- Glucose detection kit was used to detect the glucose consumption capacity of peritoneal monocytes and macrophages pre-programmed by IGF2.
- IGF2 insulin growth factor receptor 2
- Glucose Colorimetric Assay Kit II BioVision, K686-100.
- the rate of glucose consumption can reflect the type of energy metabolism and inflammatory phenotype of macrophages.
- the lysosomes of IGF2-treated peritoneal monocytes and macrophages were separated, and the experiment was performed according to the experimental method provided by the Lysosome Isolation Kit (Sigma-Aldrich, LYSISO1-1KT).
- the separated lysosomes will be used in the V-type ATPase activity detection experiment.
- Use the ATPase activity detection kit to detect the lysosomal ATPase activity isolated above, which represents the activity of lysosomal type V ATPase.
- the data are expressed in the form of mean or mean ⁇ S.E.M. or S.D., using Graphpad software for mapping and significant difference analysis.
- significance of the difference between the two sets of data was analyzed using two-tailed non-parametric Student’s t test. ns means insignificant difference; *P ⁇ 0.05; **P ⁇ 0.01; ***P ⁇ 0.001, P ⁇ 0.05 is considered a significant difference.
- Example 1 Low-dose IGF2 inhibits peritonitis.
- L-IGF2 Low-dose IGF2
- H-IGF2 high-dose IGF2
- Example 2 Anti-inflammatory macrophages under the control of low-dose IGF2 are dominated by the metabolic preference of oxidative phosphorylation.
- macrophages Under the control of low-dose IGF2, macrophages exhibit higher levels of OCR:ECAR, indicating that this group of macrophages tend to use oxidative phosphorylation (OXPHOS) as the main way to obtain energy, while the control group and high-dose IGF2 Group macrophages are more inclined to choose aerobic glycolysis to produce energy for the cells.
- OXPHOS oxidative phosphorylation
- Lactic acid is an intermediate metabolite of aerobic glycolysis. In the process of aerobic glycolysis, excess lactic acid will be secreted out of the cell by the cells. Therefore, to detect and compare the cumulative amount of lactic acid in the supernatant of the cell culture medium, it is An effective way to measure the level of aerobic glycolysis in cells.
- LPS Lipopolysaccharide
- LPS lipopolysaccharide
- IL-4/113 interleukin-4/13
- RT-PCR results showed that the expression of inflammatory factors in macrophages classically activated by lipopolysaccharide was inhibited by low-dose IGF2 (L-IGF2), and high-dose IGF2 had no significant regulation on some genes (Figure 2D); the anti-inflammatory factor expression of macrophages activated by interleukin 4/13 is amplified by IGF2 (L-IGF2), and high-dose IGF2 (H-IGF2) has no obvious effect (Figure 2E).
- L-IGF2 low-dose IGF2
- H-IGF2 high-dose IGF2
- Example 3 Low-dose IGF2 inhibits type V ATPase activity, reduces lysosomal H + concentration, promotes cytoplasmic H + concentration, and increases mitochondrial inner membrane potential.
- Oxidative phosphorylation occurs in the mitochondria, while aerobic glycolysis occurs in the cytoplasm.
- IGF2 enhances the oxidative phosphorylation capacitation preference of macrophages.
- peritoneal macrophages mature and mature macrophages treated with low-dose IGF2 (L-IGF2, 50ng per mouse) all showed higher mitochondrial membrane potential ( Figure 3A), suggesting a significant increase The mitochondrial membrane potential provides sufficient H + substrates for the oxidative phosphorylation of macrophages under the control of low-dose IGF2.
- Example 4 Construction of igf2r gene-deficient mice and igf1r or igf2r conditional knockout mice.
- IGF2 can bind to IGF1R and IGF2R, and has the greatest affinity with IGF2R. When IGF2 binds to IGF2R, it causes IGF2R to be internalized, and IGF2 will be transported to the lysosome for degradation. Therefore, IGF2R is often considered as a decoy receptor for IGF2. ), and the combination of IGF2 and IGF1R exerts mitogen function and promotes cell proliferation and survival 2 . According to the previous part of the study, we found that high-dose IGF2 reflects the function of mitogen and promotes the proliferation and survival of peritoneal macrophages in peritonitis mice, while low-dose IGF2 has no such function.
- IGF2 In view of the maximum affinity of IGF2 and IGF2R, we hypothesize that low-dose IGF2 preferentially binds to IGF2R, and induces IGF2R-dependent biological effects, such as the occurrence of non-classical mitochondrial dynamics. High-dose IGF2 “depletes” the outer cell membrane IGF2R. There is still a chance to combine with IGF1R to counteract the biological effects of IGF2R, thereby promoting the proliferation and survival of peritoneal macrophages in peritonitis mice.
- mice In order to study which receptors IGF2 relies on to achieve bidirectional regulation of monocytes, we constructed genetically engineered mice. In igf2r gene knockout mice, we used Crisper-Cas9 technology to target and knock out the 2bp or 13bp base of exon 6 of igf2 gene, causing frameshift mutation to complete ( Figure 4A). In view of the fact that complete deletion of igf2r gene can cause mouse embryonic death, we finally obtained igf2r knockout heterozygous mice-IGF2R +/- mice for breeding and conservation and experiments, and identification of IGF2R +/- mouse progeny genotypes Rely on sequencing alignment (Figure 4B). We induced mouse peritoneal macrophages, and used flow cytometry to detect the expression of IGF2R in these cells to confirm the knockout efficiency (Figure 4C).
- IGF1R fl/fl mice and IGF2R fl/fl mice were bred with Lyz2 Cre mice respectively, and Lyz2 was expressed at a high level in myeloid cells 18 until IGF1R fl/fl Lyz2 Cre homozygous mice and IGF2R fl/ fl Lyz2 Cre homozygous mice ( Figure 4D, G).
- the DNA level and protein level identification results verified the knockout efficiency ( Figure 4E, F, H, I).
- DSS dextran sulfate sodium salt
- IGF2 inhibits DSS-induced inflammatory bowel disease
- the Ki67 positive signal of the low-dose IGF2 group was significantly stronger than the control group and the high-dose IGF2 treatment group, which indicates that the basement membrane is near
- the monocytes and macrophages played a role in the repair of intestinal villi.
- the macrophages in the colonic basement membrane of mice treated with low-dose IGF2 could better promote colonic villi repair (Figure 5H).
- low-dose IGF2 can significantly improve the quality of life and survival time of DSS-induced inflammatory bowel disease mice, while high-dose IGF2 has no therapeutic effect and may even aggravate some symptoms.
- IGF2 treatment of inflammatory bowel disease relies on macrophages
- macrophages regulated by low-dose IGF2 can alleviate inflammatory bowel disease, while macrophages regulated by high-dose IGF2 aggravate the progression of inflammatory bowel disease.
- IGF1 aggravates inflammatory bowel disease
- IGF2 preferentially binds to IGF2R at low doses, and can bind to IGF1R at high doses. And IGF1 has the ability to bind IGF1R, but not IGF2R. In order to prove the regulatory effect of IGF1R activation on the inflammatory response, the use of IGF1 to treat inflammatory bowel disease was studied.
- IGF2R performs the function of low-dose IGF2 to induce anti-inflammatory macrophages
- the study uses myeloid cells to specifically knock out IGF2R mice, establishes a peritonitis model, and analyzes the presence or absence of low-dose IGF2 (L-IGF2, 50ng each (Mice) when injected, the expression levels of PD-L1, IL-1 ⁇ and H + in the cytoplasm of peritoneal macrophages.
- L-IGF2 low-dose IGF2
- IGF2R performs the anti-inflammatory task of low-dose IGF2
- IGF1R fl/fl Lyz2 Cre mice that specifically knock out IGF1R are more resistant to inflammatory bowel disease, no matter in the PBS treatment group or high-dose IGF2(H -IGF2, 1000ng per mouse) in the treatment group, the weight of the mice was significantly higher than the weight of the control group IGF1R fl/fl mice under the corresponding treatment conditions, the survival of the mice was significantly prolonged, the stool score and the degree of blood in the stool were significantly reduced, and the colon Inflammatory infiltration was significantly reduced (Figure 10G-L).
- IGF2 mutants In order to facilitate the study of the functions of IGF2R and IGF1R, many IGF2 mutants have been designed and reported, including IGF2 mutants that can selectively bind to IGF2R.
- the researchers mutated the tyrosine at position 27 of the IGF2 peptide to leucine to prepare the Leu27-IGF2 mutant (L27-IGF2).
- the Leu27-IGF2 mutant has the same affinity for IGF2R and IGFBP protein, and has the same affinity with IGF1R. The affinity is extremely reduced.
- Leu27-IGF2 can significantly reduce the relative pH of the cytoplasm of peritonitis mouse peritoneal monocytes and peritoneal macrophages. Even at a high dose of 2000ng/ml, we also compared Leu27-IGF2 at a low dose. The effect of changing the pH of the cytoplasm eliminates the possibility of the low titer of Leu27-IGF2 ( Figure 11A). In the study of peritonitis, we found that macrophages after treatment with low-dose Leu27-IGF2 and high-dose Leu27-IGF2 both showed significant production of nitric oxide ( Figure 11B) and lactic acid (Figure 11C) in response to LPS stimulation. decline.
- IGF2 in this part of the study, we verified the therapeutic effect of IGF2 in a mouse model of DSS-induced inflammatory bowel disease. It was confirmed that low-dose IGF2 effectively inhibited inflammatory bowel disease depends on the same mechanism of action as low-dose IGF2 in the treatment of peritonitis. In both validation models, IGF2R-dependent pathways can give macrophages the immune memory that significantly inhibits inflammation. IGF2 mutants target to activate IGF2R and exert the most stable effect of inhibiting inflammation.
- Human Leu27-IGF2 is an analogue of IGF-2, which replaces tyrosine with leucine at position 27 of the human IGF2 sequence. Compared with IGF2, the binding ability of Leu27-IGF2 with serum IGF-binding proteins (IGFBPs) remains unchanged, but the binding ability with IGF1 receptor (IGF1R) is reduced by 10-20 times. Therefore, Leu27-IGF2 and IGF2 receptor ( The specificity of IGF2R) binding is enhanced.
- the IGF2 peptide consists of four domains (Domains) A, B, C, and D. The deletion of the D domain (amino acids 62-67) will significantly enhance its affinity with IGFBP2, IGFBP3, IGFBP4 and IGFBP6, and with IGFBP1, The affinity of IGFBP5 remains unchanged.
- IGF2R Activation of IGF2R promotes its nuclear transport and regulates the cytoplasmic H + concentration of macrophages
- IGF2 can reduce the distribution of IGF2R on the cell membrane of THP1 cells, showing a dose-dependent characteristic. Starting from a low dose of 5ng/ml to a high dose of 1000ng ml, the reduction of IGF2R on the cell membrane surface is maximized. Continue to increase the dose of IGF2 on the cell membrane surface. IGF2R no longer continued to decrease ( Figure 13A, B). Using western blotting, it is clear that after treatment of THP1 cells with IGF2, the distribution of IGF2R in the nucleus is significantly increased ( Figure 13C). Blocking the nuclear transport of IGF2R with Ivermectin can inhibit the distribution of IGF2R in the nucleus ( Figure 13D).
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Abstract
Description
最初的残基 | 代表性的取代 | 优选的取代 |
Ala(A) | Val;Leu;Ile | Val |
Arg(R) | Lys;Gln;Asn | Lys |
Asn(N) | Gln;His;Lys;Arg | Gln |
Asp(D) | Glu | Glu |
Cys(C) | Ser | Ser |
Gln(Q) | Asn | Asn |
Glu(E) | Asp | Asp |
Gly(G) | Pro;Ala | Ala |
His(H) | Asn;Gln;Lys;Arg | Arg |
Ile(I) | Leu;Val;Met;Ala;Phe | Leu |
Leu(L) | Ile;Val;Met;Ala;Phe | Ile |
Lys(K) | Arg;Gln;Asn | Arg |
Met(M) | Leu;Phe;Ile | Leu |
Phe(F) | Leu;Val;Ile;Ala;Tyr | Leu |
Pro(P) | Ala | Ala |
Ser(S) | Thr | Thr |
Thr(T) | Ser | Ser |
Trp(W) | Tyr;Phe | Tyr |
Tyr(Y) | Trp;Phe;Thr;Ser | Phe |
Val(V) | Ile;Leu;Met;Phe;Ala | Leu |
Claims (18)
- 一种非IGF1R结合型的物质的用途,其特征在于,用于制备组合物或制剂,所述组合物或制剂用于预防和/或治疗炎症性疾病。
- 如权利要求1所述的用途,其特征在于,所述非IGF1R结合型的物质选自下组:IGF2突变体、表达IGF2突变体的载体、抗体、小分子化合物、或其组合。
- 如权利要求1所述的用途,其特征在于,所述炎症性疾病选自下组:腹膜炎、炎症性肠病、多发性硬化、糖尿病、系统性红斑狼疮、硬皮病、桥本甲状腺炎、自身免疫性肝炎、自身免疫葡萄膜炎、间质性肺病、银屑病、白癜风、皮肌炎、川崎病、成人耳氏病、强制性脊柱炎、类肉状瘤病、起止点炎相关的关节炎、多关节青少年特发性关节炎、类风湿性关节炎、移植物抗宿主病、自身免疫性胰腺炎、帕金森病、老年痴呆、或其组合。
- 如权利要求1所述的用途,其特征在于,所述非IGF1R结合型的物质包括对IGF2R的选择性或亲和力高于IGF1R的物质,以及可以高效激活IGF2R的不激活或基本不激活IGF1R的物质。
- 如权利要求2所述的用途,其特征在于,所述IGF2突变体在野生型IGF2蛋白的对应于SEQ ID NO.:1的第27位的酪氨酸发生突变。
- 如权利要求2所述的用途,其特征在于,所述IGF2突变体在野生型IGF2蛋白的对应于SEQ ID NO.:1的第27位的酪氨酸突变为亮氨酸,并任选的删除D-domain(SEQ ID NO.:1的第62位苏氨酸至第67位谷氨酸)。
- 如权利要求2所述的用途,其特征在于,所述IGF2突变体选自下组:(a)具有SEQ ID NO.:2-58任一所示氨基酸序列的多肽;(b)将SEQ ID NO.:2-58任一所示氨基酸序列经过一个或多个(如2个、3个、4个或5个)氨基酸残基的取代、缺失或添加而形成的,且具有抑制炎症活性的由(a)衍生的多肽。
- 一种细胞制剂,其特征在于,包括:经过非IGF1R结合型的物质处理的吞噬细胞。
- 如权利要求8所述的细胞制剂,其特征在于,所述吞噬细胞选自下组:单核细胞、巨噬细胞、单核细胞前体细胞、或其组合。
- 如权利要求8所述的细胞制剂,其特征在于,所述的组合物中,所述吞噬细胞的浓度1×10 4-5×10 7/ml,较佳地5×10 4-5×10 6/ml,更佳地 1×10 5-1×10 6/ml。
- 一种药盒,其特征在于,包括:(i)第一容器,以及装于该第一容器中的活性成分(a)经过非IGF1R结合型的物质处理的吞噬细胞,或含有活性成分(a)的药物;(ii)任选的第二容器,以及装于该第二容器中的活性成分(b)非IGF1R结合型的物质,或含有活性成分(b)的药物;和(iii)说明书,所述说明书中记载了联合给予活性成分(a)和活性成分(b)从而预防和/或治疗炎症性疾病的说明。
- 一种筛药方法,其特征在于,包括步骤:(a)测试组中,在细胞的培养体系中添加测试物质,并观察所述测试组的细胞中所述测试物质与IGF1R和/或IGF2R的结合活性;在对照组中,在相同细胞的培养体系中不添加测试物质;其中,如果测试组的细胞中所述测试物质与IGF1R的结合活性降低;并且与IGF2R的结合活性不变或升高,则表明,所述测试物质为非IGF1R结合型的IGF2突变型。
- 一种筛选预防和/或治疗炎症性疾病的候选物的方法,其特征在于,包括:(a)测试组中,在细胞的培养体系中添加测试物质,并观察所述测试组的细胞中所述测试物质对IGF2R在细胞表面的表达和/或活性的影响;和/或所述测试物质对IGF2R核转运的影响;和/或所述测试物质对IGF2R向细胞核的重分布的影响;在对照组中,在相同细胞的培养体系中不添加测试物质;其中,如果测试组的所述测试物质抑制IGF2R在细胞表面的表达和/或活性;和/或所述测试物质增加IGF2R的核转运;和/或所述测试物质促进IGF2R向细胞核的重分布,则表明,所述测试物质为预防和/或治疗炎症性疾病的候选物。
- 一种筛药方法(或筛选候选物的方法),其特征在于,包括:(a)测试组中,在细胞的培养体系中添加测试物质,并观察所述测试组的细胞中所述测试物质对IGF2与IGF1R和/或IGF2R的结合活性的影响;在对照组中,在相同细胞的培养体系中不添加测试物质;其中,如果测试组的细胞中所述测试物质抑制IGF2与IGF1R的结合活性;并且对IGF2与IGF2R的结合活性无影响或有促进作用,则表明,所述测试物质为非IGF1R结合型的物质(或候选物)。
- 一种体外获得促炎样本和抑炎样本的方法,其特征在于,包括:在第一组实验中,用低剂量的IGF2进行处理,获得抑炎样本;和在第二组实验中,用高剂量的IGF2进行处理,获得促炎样本。
- 一种组合物,其特征在于,包括:(a)权利要求12或13或14所述的方法获得的非IGF1R结合型的IGF2突变型或候选物;其中所述非IGF1R结合型的IGF2突变型不包括选自下组的IGF2突变型:野生型IGF2、野生型IGF2第25~91位氨基酸片段、12位谷氨酸突变为Asp、Ala、Gln、His、Arg、或Lys的IGF2突变型、26位苯丙氨酸突变为Ser的IGF2突变型、27位酪氨酸突变为Leu的IGF2突变型、43位缬氨酸突变为Leu的IGF2突变型、删除D-domain的IGF2突变型、或其组合。
- 一种抑制炎症活性的方法,其特征在于,包括步骤:在非IGF1R结合型的物质(或具有IGF2R选择性的候选物)存在下,培养吞噬细胞,从而抑制炎症活性。
- 一种权利要求8所述的细胞制剂、权利要求11所述的药盒或权利要求16所述的组合物的用途,其特征在于,用于制备用于预防和/或治疗炎症性疾病的药物。
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AU2020346948A AU2020346948A1 (en) | 2019-09-10 | 2020-09-10 | Application of non-IGF1R-binding substance in prevention and/or treatment of inflammatory diseases |
CN202080006025.6A CN113164606A (zh) | 2019-09-10 | 2020-09-10 | 非igf1r结合型的物质在预防和/或治疗炎症性疾病中的应用 |
EP20863556.5A EP4046658A4 (en) | 2019-09-10 | 2020-09-10 | APPLICATIONS OF IGF1R-FREE BINDING SUBSTANCE IN THE PREVENTION AND/OR TREATMENT OF INFLAMMATORY DISEASES |
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090275608A1 (en) * | 2008-02-04 | 2009-11-05 | Bipar Sciences, Inc. | Methods of diagnosing and treating parp-mediated diseases |
WO2011098400A1 (en) * | 2010-02-11 | 2011-08-18 | F. Hoffmann-La Roche Ag | Igf-i poly (ethylene glycol) conjugates |
CN106167789A (zh) * | 2015-05-21 | 2016-11-30 | 中国科学院上海生命科学研究院 | 低氧处理的间充质干细胞及其应用 |
CN107582566A (zh) * | 2016-07-07 | 2018-01-16 | 中国科学院上海生命科学研究院 | 通过多胺化合物调控自身免疫性疾病的方法和组合物 |
WO2018099363A1 (zh) * | 2016-11-29 | 2018-06-07 | 中国科学院上海生命科学研究院 | 含胰岛素样生长因子-2的药物组合物及其应用 |
CN109053875A (zh) * | 2018-08-31 | 2018-12-21 | 重庆大学 | 突变型igf-1、重组质粒、重组蛋白及应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB201721287D0 (en) * | 2017-12-19 | 2018-01-31 | Repos Pharma Ab | Treatment for inflammatory disease |
-
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- 2020-09-10 CN CN202080006025.6A patent/CN113164606A/zh active Pending
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Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090275608A1 (en) * | 2008-02-04 | 2009-11-05 | Bipar Sciences, Inc. | Methods of diagnosing and treating parp-mediated diseases |
WO2011098400A1 (en) * | 2010-02-11 | 2011-08-18 | F. Hoffmann-La Roche Ag | Igf-i poly (ethylene glycol) conjugates |
CN106167789A (zh) * | 2015-05-21 | 2016-11-30 | 中国科学院上海生命科学研究院 | 低氧处理的间充质干细胞及其应用 |
CN107582566A (zh) * | 2016-07-07 | 2018-01-16 | 中国科学院上海生命科学研究院 | 通过多胺化合物调控自身免疫性疾病的方法和组合物 |
WO2018099363A1 (zh) * | 2016-11-29 | 2018-06-07 | 中国科学院上海生命科学研究院 | 含胰岛素样生长因子-2的药物组合物及其应用 |
CN109053875A (zh) * | 2018-08-31 | 2018-12-21 | 重庆大学 | 突变型igf-1、重组质粒、重组蛋白及应用 |
Non-Patent Citations (25)
Title |
---|
A.H.A. A. B. L.: "Basic Immunology. Functions and disorders of the immune system", 2009, ELSEVIER, article "Ch.2 Innate Immunity" |
BORGELT, L. M.: "Immunity and autoimmune diseases", WOMEN'S HEALTH ACROSS THE LIFESPAN: A PHARMACOTHERAPEUTIC APPROACH, 2010 |
CHASSAING, B.AITKEN, J. D.MALLESHAPPA, MVIJAY-KUMAR, M: "Dextran sulfate sodium (DSS)-induced colitis in mice", CURRENT PROTOCOLS IN IMMUNOLOGY, vol. 104, 2014, XP055723883, DOI: 10.1002/0471142735.im1525s104 |
CLAUSEN, B. E.BURKHARDT, C.REITH, W.RENKAWITZ, RFORSTER, I: "Conditional gene targeting in macrophages and granulocytes using LysMcre mice", TRANSGENIC RESEARCH, 1999 |
COSTELLO, M.BAXTER, R. C.SCOTT, C. D.: "Regulation of soluble insulin-like growth factor II/mannose 6-phosphate receptor in human serum: measurement by enzyme-linked immunosorbent assay", THE JOURNAL OF CLINICAL ENDOCRINOLOGY AND METABOLISM, vol. 84, 1999, pages 611 - 617 |
DU LIMING, LIN LIANGYU, LI QING, LIU KELI, HUANG YIN, WANG XUEFENG, CAO KAI, CHEN XIAODONG, CAO WEI, LI FENGYING, SHAO CHANGSHUN, : "IGF-2 Preprograms Maturing Macrophages to Acquire Oxidative Phosphorylation-Dependent Anti-inflammatory Properties", CELL METABOLISM, CELL PRESS, UNITED STATES, vol. 29, no. 6, 1 June 2019 (2019-06-01), United States, pages 1363 - 1375.e8, XP055790728, ISSN: 1550-4131, DOI: 10.1016/j.cmet.2019.01.006 * |
ENGUITA-GERMAN, MFORTES, P: "Targeting the insulin-like growth factor pathway in hepatocellular carcinoma", WORLD JOURNAL OF HEPATOLOGY, vol. 6, 2014, pages 716 - 737 |
FERRERO-MILIANI, L.NIELSEN, O. H.ANDERSEN, P. S.GIRARDIN, S. E.: "Chronic inflammation: importance of NOD2 and NALP3 in interleukin-lbeta generation", CLINICAL AND EXPERIMENTAL IMMUNOLOGY, vol. 147, 2007, pages 227 - 235 |
GARY-BOBO, M.NIRDE, P.JEANJEAN, A.MORERE, AGARCIA, M: "Mannose 6-phosphate receptor targeting and its applications in human diseases", CURRENT MEDICINAL CHEMISTRY, vol. 14, 2007, pages 2945 - 2953, XP002557212, DOI: 10.2174/092986707782794005 |
HANAYAMA RT. M.MIWA KSHINOHARA AIWAMATSU ANAGATA S: "Identification of a factor that links apoptotic cells to phagocytes", NATURE, vol. 417, no. 6885, 2002, pages 182 - 7, XP002965210, DOI: 10.1038/417182a |
HOCKING, P. M.ROBERTSON, G. W.GENTLE, M. J.: "Effects of anti-inflammatory steroid drugs on pain coping behaviours in a model of articular pain in the domestic fowl", RESEARCH IN VETERINARY SCIENCE, vol. 71, 2001, pages 161 - 166 |
KILLIAN, J. K.JIRTLE, R. L.: "Genomic structure of the human M6P/IGF2 receptor", MAMMALIAN GENOME : OFFICIAL JOURNAL OF THE INTERNATIONAL MAMMALIAN GENOME SOCIETY, vol. 10, 1999, pages 74 - 77 |
KUMEJI TAKEUCHI, N. I.NOBORU HASEGAWAYOKO YOSHIDAKENJI YAMADAHIROSHI KOGOYOSHIO AIZAWA: "Inhibitory action of non-steroidal anti-inflammatory drugs on prostaglandin synthesis and release", FOLIA PHARMACOLOGICA JAPONICA, vol. 76, 1980, pages 525 - 531 |
MARGOT W BEUKERS, YOUNGMAN OH, HEPING ZHANG, NICHOLAS LING, RON G ROSENFELD: "[Leu27] INSULIN-LIKE GROWTH FACTOR II IS HIGHLY SELECTIVE FOR THE TYPE-II IGF RECEPTOR IN BINDING, CROSS-LINKING AND THYMIDINE INCORPORATION EXPERIMENTS", ENDOCRINOLOGY, vol. 128, no. 2, 1 February 1991 (1991-02-01), US , pages 1201 - 1203, XP009535048, ISSN: 0013-7227, DOI: 10.1210/endo-128-2-1201 * |
MENKES, C. J.: "Effects of disease-modifying anti-rheumatic drugs, steroids and non-steroidal anti-inflammatory drugs on acute-phase proteins in rheumatoid arthritis", BRITISH JOURNAL OF RHEUMATOLOGY, vol. 3, 1993, pages 14 - 18 |
MIKSA, M ET AL.: "Maturation-induced down-regulation of MFG-E8 impairs apoptotic cell clearance and enhances endotoxin response", INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, vol. 22, 2008, pages 743 - 748 |
MIKSA, M.AMIN, D.WU, R.RAVIKUMAR, T. S.WANG, P: "Fractalkine-induced MFG-E8 leads to enhanced apoptotic cell clearance by macrophages", MOLECULAR MEDICINE, vol. 13, pages 553 - 560 |
OHUCHI, S. T. S. S.: "Inflammation: Mechanisms and Treatment. Inflammation: Mechanisms and Treatment", vol. 4, 1980, WILLOUGHBY D.A., article "A review of mechanism of action of steroid and non-steroid anti-inflammatory drugs" |
ROTH, B.V. ; BURGISSER, D.M. ; LUTHI, C. ; HUMBEL, R.E.: "Mutants of human insulin-like growth factor II: Expression and characterization of analogs with a substitution of Tyr^2^7 and/or a deletion of residues 62-67", BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ELSEVIER, AMSTERDAM NL, vol. 181, no. 2, 16 December 1991 (1991-12-16), Amsterdam NL, pages 907 - 914, XP024840430, ISSN: 0006-291X, DOI: 10.1016/0006-291X(91)91277-J * |
See also references of EP4046658A4 |
SHEET, A: "d. f.", 2016, DEPARTMENT OF HEALTH AND HUMAN SERVICES |
WANG, ZHONGYAN : "Research Advances in Insulin-like Growth Factor", LIVESTOCK AND POULTRY INDUSTRY, no. 11, 31 December 2004 (2004-12-31), CN, pages 62 - 64, XP009535440, ISSN: 1008-0414, DOI: 10.19567/j.cnki.1008-0414.2004.11.040 * |
WARDEN, S. J.: "Prophylactic use of NSAIDs by athletes: a risk/benefit assessment", THE PHYSICIAN AND SPORTSMEDICINE, vol. 38, 2010, pages 132 - 138 |
YU, HROHAN, T: "Role of the insulin-like growth factor family in cancer development and progression", JOURNAL OF THE NATIONAL CANCER INSTITUTE, vol. 92, 2000, pages 1472 - 1489, XP002414394, DOI: 10.1093/jnci/92.18.1472 |
ZIGMOND, E ET AL.: "Ly6C hi monocytes in the inflamed colon give rise to proinflammatory effector cells and migratory antigen-presenting cells", IMMUNITY, vol. 37, 2012, pages 1076 - 1090 |
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AU2020346948A1 (en) | 2022-04-28 |
EP4046658A1 (en) | 2022-08-24 |
CN112546228A (zh) | 2021-03-26 |
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