41 results about "Human insulin" patented technology

The invention discloses a crystallization method of recombinant human insulin, which belongs to the technical field of biological medicine, and comprises the following steps: taking a sample containing recombinant human insulin, carrying out column chromatography salt conversion to obtain a salt conversion eluent, then adding purified water, adjusting the pH value to prepare a solution to be crystallized, and adding an acid solution, a zinc salt solution and arginine to obtain a crystallization solution; and adjusting the pH value and temperature of the crystallization liquid, and crystallizing to obtain the hexahedral crystal. According to the method, the to-be-crystallized solution is prepared from the purified and prepared qualified eluent containing the human insulin in a chromatography salt conversion mode, and the preparation mode of the to-be-crystallized solution is simple, efficient and easy to operate; the crystallization liquid system is simple, only acid, zinc salt and arginine need to be added into the to-be-crystallized solution, phenols and organic solvents do not need to be added, and chemical substance residues are avoided. The residual salt and the organic solvent in the salt conversion concentrated solution are fully utilized to participate in the crystallization reaction, the solvent does not need to be repeatedly added, the reagent is saved, and the process time is shortened.

The invention relates to the field of medical detection, in particular to a mass spectrometric detection method for enriching serum human insulin by using specific antibody immunomagnetic beads. According to the present invention, the human insulin in the serum is subjected to immune enrichment by using the magnetic beads coated with the insulin specific antibody, and the concentration level of the human insulin in the serum is accurately determined based on the accurate quantification technology of the isotope dilution liquid chromatography-tandem mass spectrometry platform in combination with the bracket method quantification mode. The pretreatment method provided by the invention is simple and convenient to operate, the absolute recovery rate is excellent, the sensitivity requirement is met, and the mass spectrometry method is an isotope diluent liquid chromatography-tandem mass spectrometry method capable of eliminating interference of structural analogues.

The invention discloses an NK cell serum-free medium, the serum-free medium comprises a basic medium and an additive, the additive comprises the following components: recombinant human transferrin, recombinant human insulin, human serum albumin, growth factors, glucose, trace elements, tacrolimus and mevalonic acid, the serum-free culture medium comprises the following components in percentage by weight: 85% of a basic culture medium, 3% of recombinant human transferrin, 3% of recombinant human insulin, 2% of human serum albumin, 2% of growth factors, 2% of glucose, 1.5% of trace elements, 1% of tacrolimus and 0.5% of mevalonic acid; according to the serum-free culture medium for the NK cells, growth and differentiation of the NK cells can be adjusted by adding a proper amount of tacrolimus, meanwhile, a proper amount of mevalonic acid is matched, raw materials of active substances can be provided for the cultured NK cells, growth and development of the NK cells or expression of specific functions are facilitated, and therefore the survival rate and the amplification number of the NK cells can be increased.

The invention discloses a serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and application thereof. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition is prepared from the following components in parts by mass: 15 to 25 parts of platelet-derived growth factor-BB; 15 to 25 parts of basic fibroblast growth factors; 8 to 12 parts of transforming growth factor-beta; and 8-12 parts of recombinant human insulin. The serum substitute compatible umbilical cord mesenchymal stem cell culture medium additive composition and the application thereof disclosed by the invention have the following beneficial effects: (1) the proliferation efficiency is remarkably improved; (2) the cell morphology and dryness can be well maintained; and (3) the compliance and the compatibility are good.

The invention provides a high-stability high-concentration zinc-containing insulin preparation, and belongs to the field of pharmaceutical preparations, the concentration of human insulin in the preparation is greater than 500 IU / mL, the preparation further contains zinc ions, a preservative, an isotonic agent and a buffer agent, the pH value of the preparation is 7.0-7.8, and the content of the zinc ions in the preparation is greater than 1%. Compared with similar products known in the field, the preparation has great physical stability, the volume of injection liquid medicine can be reduced under the condition that the required amount of insulin is the same, the hardening and caking phenomena of tissues at an injection part are effectively relieved, and the preparation still can be kept in a basically non-aggregated state in a heat or mechanical pressure environment; the method is suitable for continuous infusion systems.

The invention relates to the technical field of biology, and discloses a culture medium system for pluripotency maintenance and directional induction of embryonic stem cells and application of the culture medium system. 0.5 to 0.8 part of tetramethylenediamine; 0.2 to 0.5 part of corpus luteum hormone; 1-2 parts of a vitamin mixture; 2-4 parts of bovine serum albumin; 0.5 to 1.0 part of sodium bicarbonate; 0.2 to 0.4 part of an antibiotic mixed solution; 1 to 2 parts of biotin; 0.5 to 1.0 part of bone morphogenetic protein; and 0.1 to 0.2 part of magnesium ethylene diamine tetraacetate. A multi-factor regulation and control system is constructed, signal inhibition, a three-dimensional scaffold and a metabolic regulation module are combined, pluripotency of the embryonic stem cells is stably maintained, culture uniformity, structural support and differentiation resistance are remarkably improved, and the technical bottleneck of a traditional system in the aspects of induction efficiency and stability is broken through.

The invention provides an induced pluripotent stem cell culture medium and a culture method of induced pluripotent stem cells. The induced pluripotent stem cell culture medium comprises a basic culture medium, a composite additive A and a composite additive B, the composite additive A comprises 5-30 [mu] g / mL of recombinant human insulin growth factors, 1-5 ng / mL of recombinant human transforming growth factors, 50-200 ng / mL of recombinant human basic fibroblast growth factors, 7-20 [mu] g / mL of sodium selenite, 0.5-2 mg / mL of albumin, 50-70 [mu] g / mL of ascorbic acid and 5-20 [mu] g / mL of transferrin; the composite additive B is prepared from 50 to 300mg / mL of a Stock B solution, 0.5 to 5 percent (v / v) of a non-essential amino acid solution, 50 to 200ng / mL of DL-piperidinecarboxylic acid, 50 to 200mu g / mL of gamma-aminobutyric acid, 0.1 to 1 percent (v / v) of a lipid concentrated solution, 0.5 to 5 percent (v / v) of an L-alanyl-L-glutamine solution and 0.1 to 0.1 percent of mercaptoethanol. The culture medium for the induced pluripotent stem cells provided by the invention can ensure the self-renewal capability of the iPSCs in a long-term passage process; and the genetic stability of the iPSCs cells is optimized according to the genome integrity and pluripotent state.

The present disclosure describes a human umbilical cord mesenchymal stem cell culture medium and a preparation method thereof, wherein the human umbilical cord mesenchymal stem cell culture medium includes a basal culture medium, reduced glutathione, epidermal growth factor, basic fibroblast growth factor, recombinant human insulin, hyaluronic acid and fibronectin, and the human umbilical cord mesenchymal stem cell culture medium does not include animal serum, and the reduced glutathione concentration is 1 mg / L to 3 mg / L, the epidermal growth factor concentration is 30 μg / L to 80 μg / L, the basic fibroblast growth factor concentration is 10 μg / L to 30 μg / L, the recombinant human insulin concentration is 1 mg / L to 5 mg / L, the hyaluronic acid mass fraction is 0.1% to 0.6%, and the fibronectin concentration is 10 mg / L to 50 mg / L. According to the present disclosure, a human umbilical cord mesenchymal stem cell culture medium and a preparation method thereof that does not contain animal serum, can support primary cell culture, is conducive to cell proliferation in vitro, adheres to the wall and maintains stemness can be provided.

The invention belongs to the technical field of immune cell culture, and particularly relates to an NK cell in-vitro amplification culture medium and a culture method thereof. The culture medium consists of a basic culture medium and an auxiliary additive, wherein the basic culture medium is an X-VIVO 15 culture medium; the auxiliary additive is prepared from the following components: human serum albumin HSA, IL-12, IL-18, IL-15, rapamycin, nicotinamide, glute-IL-15 microspheres, alginate-IL-21 microspheres, recombinant human insulin, transferrin, glutamine, beta-mercaptoethanol, non-essential amino acid and a lipid mixture. According to the invention, efficient amplification and long-acting cytotoxicity are realized through sequential synergy of IL-15 slow release / IL-21 pulse release and metabolism synergy of NAD < + >-mitochondria / lipid-membrane homeostasis.

Polypeptide antagonists of the human insulin receptor are provided. In some embodiments, a pharmaceutical formulation is provided, comprising an INSR antagonist as identified herein, and a pharmaceutically acceptable excipient. The formulation may be provided in a unit dose, e.g. a therapeutically effective dose. In methods of the disclosure, an effective dose of a an INSR antagonist as identified herein is administered to an individual having, or at risk of having, hyperinsulinemia, in a dose effective to stabilize, reduce or prevent clinical symptoms of the disease. The individual may be monitored for clinical indicia of disease before, during, and / or after administration.

The application discloses a preparation method of a human insulin purity standard substance, which comprises the following steps of raw material selection, qualitative identification, standard substance uniformity, stability test, value determination and uncertainty evaluation. The application uses commercially available high-purity human insulin as a raw material, and after purity analysis, qualitative identification, uniformity test and stability test, isotope-labeled Val, Leu and Phe are used as internal standards, and liquid chromatography-isotope dilution mass spectrometry is used for value determination, so that the interference and influence of possible isoleucine substitution leucine impurities, isomer impurities or other molecular weight protein impurities on the value determination result can be effectively excluded, and the value determination result is more accurate and reliable. The purity standard substance prepared by the application can be used for serum insulin standard substance quality value tracing, quality control of insulin measurement results in in-vitro diagnosis, related method confirmation and evaluation, and evaluation of repeatability, linearity and cross contamination rate of a closed light-emitting immune analyzer.

A pharmaceutical formulation presented, comprising an insulin or an insulin analog, and at least one protective excipient, in high concentration, selected from: a protective osmolyte, a surfactant, and a phenolic preservative. In embodiments, the insulin or insulin analog is one or more of: regular human insulin, insulin lispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, or insulin degludec. In some embodiments, at least one of: the concentration of the protective osmolyte confers an osmolarity greater than 250 mOsm / L, the concentration of the surfactant is greater than 0.1% w / v, or the concentration of the phenolic preservative is greater than 0.4% w / v. In some embodiments, insulin API is dissolved and incubated for 20 to 40 minutes at high pH before the protective excipients are added.

The invention discloses a composition for treating diabetes mellitus. The composition is prepared from the following active ingredients in parts by weight: 15-25 parts of dietary fiber powder, 3-8 parts of brown algae powder, 2-5 parts of aloe vera gel powder, 2-5 parts of green coffee powder, 1-4 parts of agaricus bisporus concentrated powder, 1-3 parts of mulberry leaf powder, 1-3 parts of rhizoma polygonati powder, 1-3 parts of bitter gourd powder and 1-2 parts of fructus momordicae powder. The dietary fiber powder and the agaricus bisporus concentrated powder supplement dietary fibers and effectively delay absorption, meanwhile, plant active ingredients in the brown algae powder, the green coffee powder and the mulberry leaf powder inhibit digestive enzymes, digestion and absorption of carbon water are blocked from the source, double barriers are achieved, and blood sugar is effectively controlled; the aloe vera gel powder and the rhizoma polygonati powder improve the insulin sensitivity and improve the insulin resistance; in addition, the balsam pear powder simulates the insulin function of a human body, and the momordica grosvenori powder reduces the sugar increasing load while adjusting the taste, so that the stability of blood sugar is maintained together.

This invention belongs to the field of stem cell culture medium technology, and provides a cell culture medium for enriching extracellular vesicles derived from stem cells and its preparation method. This invention disperses sodium hyaluronate or sodium hyaluronate-chondroitin sulfate sodium on a defined basic culture medium and grafts porous calcium alginate-gelatin microcarriers onto its surface, while limiting the content of recombinant human insulin, recombinant human transferrin, sodium selenite, and recombinant human serum albumin. This forms a culture system that combines a cell interaction interface with separable microcarriers, improving the enrichment stability of extracellular vesicles derived from stem cells, reducing the risk of solid-liquid separation residues and coupling agent residues, and solving the problem of balancing enrichment capacity with solid-liquid separation and grafting stability with biosafety. It is suitable for in vitro stem cell culture and extracellular vesicle preparation.

The application discloses a kind of serum-free epidermal melanocyte culture medium and its application, method includes by dissolving adenine, hepatocyte growth factor (HGF), glutamine, retinoic acid, vitamin E, nicotinic acid, sodium selenite, sodium hyaluronate and recombination human type XVII collagen are configured into initial culture medium;Preparation of melanocyte stimulating hormone, endothelin-3, transferrin, triiodothyronine, cholesterol, neuroregulin 1 (NRG1), 3-isobutyl-1-methyl purine (IBMX), dibutyryl cyclic adenosine monophosphate (db-cAMP), cholera toxin, hydrocortisone and recombination human insulin mother liquor;Step three: according to culture and concentration, a certain amount of mother liquor of step two is added to initial culture medium.The culture medium can effectively promote human epidermal melanocyte proliferation, melanin accumulation and help to promote human umbilical cord mesenchymal stem cells into human epidermal melanocyte, provide key advantage for obtaining functional melanocyte with high activity and therapeutic potential.

Human insulin analogs, fusion proteins and pharmaceutical uses thereof. Human insulin analog-Fc fusion proteins have a very long in vivo pharmacokinetic profile and can be used to treat metabolic diseases (e.g., type 1 diabetes and type 2 diabetes).

The invention discloses a method for improving killing activity of NK cells, and belongs to the technical field of cellular immunity. Carrying out segmented in-vitro culture on the separated NK cells by adopting a specific basic induction culture medium, an activation induction culture medium and an amplification culture medium, wherein recombinant human insulin and sodium selenite are added into the basic induction culture medium; ganoderma lucidum polysaccharides, eicosapentaenoic acid, IL-15 and IL-18 are added into the activation and induction culture medium. Salidroside, tea polyphenol, low molecular weight chitosan, IL-2 (interleukin-2), IL-21 (interleukin-21) and SCF (mesenchymal stem cells) are added into the amplification culture medium. According to the method, the killing ability of the NK cells obtained through in-vitro culture on tumor cells and the in-vitro proliferation effect of the NK cells are remarkably improved, the problems that in the prior art, the in-vitro proliferation efficiency of the NK cells is low, and cell activity is difficult to consider are solved, and a foundation is laid for clinical transformation of NK cell immunotherapy.

The invention relates to the technical field of insulin biological detection, and discloses an in vitro activity determination method for human insulin and analogues based on mouse myoblasts C2C12, which comprises the following steps: adding human insulin DMEM-H to domesticate C2C12 cells, inducing and differentiating the C2C12 cells into myotubes, and detecting the activity of the myotubes. Then adding human insulin reference products and human insulin and analogue test products with different concentration gradients to incubate myotubes, and calculating the in-vitro biological activity of the human insulin and analogue test products by comparing glucose consumption and utilization rates and carrying out parameter fitting after incubation is finished. Compared with an existing insulin activity detection method which is long in period, high in difficulty and poor in repeatability, the method in the scheme has the advantages that the sensitivity of cells to insulin is effectively improved through domestication, and the detection period is greatly shortened; and compared with a primary adipose cell method, the operation difficulty is reduced, the repeatability is better, and the method is conveniently popularized and applied to detection of the biological activity of various insulin and analogues.

Novel derivatives of human insulin are described that have binding affinities for the IGF-1 receptor surprisingly comparable to human IGF-1, while having higher affinities for both isoforms of the insulin receptor than human insulin. The novel insulin derivatives have D-histidine or glycine at position B24, the B chain has glycine at the C-terminus at position B31 and tyrosine at position B32, and glutamic or aspartic acid at position B10. The derivatives of the invention can be used particularly for growth and proliferation stimulation in cell cultures or for treatment of neurological disorders as of Alzheimer's disease, Huntington's and Parkinson's diseases, a cognitive loss or symptoms of syndromic autism, since one derivative could mediate both insulin and IGF-1 function.

Provided are a differentiation-inducing culture medium for the transformation of adipose-derived mesenchymal stem cells into chondrocytes and the use thereof. A basal culture medium of the differentiation-inducing culture medium comprises a nutrient solution and an inducer. The nutrient solution comprises a blood substitute, a non-essential amino acid, β-mercaptoethanol, penicillin / streptomycin, dexamethasone, sodium pyruvate, ascorbic acid, sodium selenite, transferrin, recombinant human insulin, β-glycerophosphate and TGF-β1; and the inducer comprises cyanamide, delphinidin chloride and an extracellular matrix derived from adipose-derived mesenchymal stem cells. The components of the inducer, i.e. cyanamide and delphinidin chloride, can stimulate chondrocyte generation by means of up-regulating the expression of aggrecan and collagen II; and the component of the inducer, i.e. extracellular matrix (ADSC-ECM), can increase the adhesion between chondrocytes, enhance the signaling communication between chondrocytes, and promote the growth and metabolism of chondrocytes, thereby greatly improving the differentiation efficiency of adipose-derived mesenchymal stem cells into chondrocytes.

The application relates to an aqueous pharmaceutical formulation comprising 200-1000 U / mL [equimolar to 200-1000 IU human insulin] of insulin glargine.

The invention concerns a soluble insulin preparation comprising NϵB29-hexadecandioyl-γ-Glu-(desB30) human insulin in a concentration from 1800 nmol / ml to 4200 nmol / ml, Zinc ions in a concentration from 4.0 Zn / 6Ins to 7.0 Zn / 6Ins, Niacinamide in a concentration from 110 mM to 220 mM or treprostinil in a concentration from 0.02 μg / ml to 1 μg / ml, Citrate in a concentration from 6 mM to 40 mM, and pH in the range from 7.0 to 8.0. The invention further comprises a method for using the soluble insulin preparation for reducing the blood glucose level in a mammal and a process for preparing the soluble insulin preparations.

The invention relates to the technical field of insulin derivatives, in particular to an insulin glycosylation derivative as well as a pharmaceutical composition and application thereof. The insulin glycosylation derivative disclosed by the invention is formed by connecting sugar with hydroxyl of human insulin B chain 27-site threonine. The insulin glycosylation derivative disclosed by the invention has higher anti-auto-agglutination tendency and solubility than insulin, also maintains the hypoglycemic activity equivalent to that of insulin, and has the potential of being developed into quick-acting insulin which takes effect more quickly.

The present disclosure relates to a composition for increasing the expression of a growth factor gene, which contains core-shell structured microparticles as an active ingredient. When administered in vivo along with a growth factor gene, the composition for increasing the expression of a growth factor gene of the present disclosure can increase the expression of the co-administered gene by at least 30%. Especially, when administered along with at least one gene selected from a human hepatocyte growth factor (HGF) gene, an isoform gene of the human hepatocyte growth factor and a variant gene thereof, or at least one gene selected from a human insulin-like growth factor 1 (IGF1) gene, an isoform gene of the human insulin-like growth factor 1 and a variant gene thereof, which are appropriate for the present disclosure, the composition can increase the expression of the gene by at least 30%. When administered along with a gene therapeutic agent, the composition can achieve a therapeutic effect even with a very small amount of a gene, and thus is useful.

The invention provides a DC-CIK cell culture medium. The DC-CIK cell culture medium comprises a DC culture medium, a DC maturation factor, a CIK culture medium, a CIK activating factor and a coating solution. The DC culture medium is prepared from IL-4, GM-CSF, recombinant human albumin, recombinant human transferrin, recombinant human insulin, cholesterol, copper sulfate, manganese chloride, ethanolamine, progesterone, palmitic acid, astragalus polysaccharide and DMEM / F12; the DC maturation factor comprises TNF-alpha, IL-1beta and a DMEM / F12 culture medium; the CIK culture medium is prepared from IMDM, lysine, polysaccharide nucleic acid, L-alanyl-L-glutamine, adenosine deaminase and glucose; the CIK activating factors comprise Anti-CD3, TNF (Tumor Necrosis Factor)-alpha, IL-1beta and DMEM (Dulbecco Modified Eagle Medium) / F12; the value-added rate and the cell viability of the DC-CIK cells cultured by the cell culture medium disclosed by the invention are remarkably improved.

The present invention provides glucose-sensitive insulin derivatives comprising a macrocycle, a glucose mimetic, and human insulin or an analogue thereof, and their pharmaceutical uses. In addition, the present invention relates to pharmaceutical compositions comprising such glucose-sensitive insulin derivatives, and to the use of such compounds in the treatment or prevention of medical conditions associated with diabetes.

The invention provides a monoclonal antibody for identifying proinsulin. A light chain CDR3 of the monoclonal antibody is shown as SEQ ID NO: 13, a light chain CDR1 of the monoclonal antibody is shown as SEQ ID NO: 14, a light chain CDR2 of the monoclonal antibody is shown as SEQ ID NO: 15, a heavy chain CDR3 of the monoclonal antibody is shown as SEQ ID NO: 16, a heavy chain CDR1 of the monoclonal antibody is shown as SEQ ID NO: 17, and a heavy chain CDR2 of the monoclonal antibody is shown as SEQ ID NO: 18. The monoclonal antibody is used for identifying human proinsulin. The monoclonal antibody can specifically recognize the specific epitope of the proinsulin, has the advantages of high recognition specificity, good affinity, high titer and the like, and has wide application value.

A method and vectors for controlling blood glucose levels in a mammal are disclosed. In one embodiment, the method comprises the steps of: treating the hepatocyte cells of a patient with a first, second or third vector, wherein the first vector comprises a promoter enhancer, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase and an albumin 3′UTR and lacks an HGH intron, wherein the second vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site and an albumin 3′UTR and lacks a promoter enhancer, wherein the third vector comprises an HGH intron, glucose inducible regulatory elements, a liver-specific promoter, a gene encoding human insulin with modified peptidase site, an albumin 3′UTR and a promoter enhancer and observing the patient's insulin levels, wherein the patient's insulin levels are controlled.

The present invention is generally directed to methods for measuring Insulin Resistance (IR) and for modulation of Type 2 Diabetes (T2D) onset treatment. In particular, diagnosis of IR is based on detection of different forms of disulfide bonds pairs that may form in insulin, affecting the normal insulin structure. Abnormal insulin is defined as any insulin that contains at least one of the forms of “abnormal” disulfide bond pairs. The methods herein measure human insulin isomers' concentration in a sample by utilizing HPLC or a monoclonal / polyclonal antibody to human insulin that contains “normal” or one or more “abnormal” disulfide bonds combinations. Some aspects of the invention also provide kits adapted for use in such methods. The human insulin isomers' concentration results are fed into a system to regulate the level of exogenous insulin needed to augment the level of normal insulin detected by the methods for a specific patient, or effectively regulate the amount of metformin, sulfonylureas, thiazolidinediones (PPARy agonists; glitazones), GLP-1s, SGLT-2 inhibitors and DPP-4 inhibitors or other IR treatment to be administered to the patient. The system and methods comprise a feedback loop to maintain a therapeutic level of insulin, metformin or other IR treatment and decrease the risk of hypoglycemia and other complications. The current invention has the advantage of being a self-referencing IR assessment by utilizing the measurements of “normal” insulin compared to “abnormal” insulin and not relying only on glucose measurements, Body Mass Index or other references included in current IR measurement methods. This leads to a much more responsive and faster treatment for IR compared to current approaches, which employ methods such as, Oral Glucose Tolerance Test (OGTT), which takes hours to perform.

The present invention provides novel salts and crystalline forms of Linsitinib (OSI‑906; cis-3-[8-amino-1-(2-phenyl-7-quinolinyl)imidazo[1,5-a]pyrazin-3-yl]-1-methyl-cyclobutanol). These new forms of Linsitinib provide increased solubility and improved pharmacokinetic profiles for orally administered pharmaceutical formulations. Accordingly, the invention enables improved methods for treating human insulin-like growth factor-1 receptor (IGF-1R) and insulin receptor (IR) mediated conditions.