50 results about "Genotoxicity" patented technology

The invention relates to a method for detecting genotoxic impurities of rupatadine fumarate, which is characterized in that a phosphate buffer solution is matched with acetonitrile, a high performance liquid chromatography method is adopted, a chromatographic column is a silica gel bonded octadecylsilane column, a gradient program is adopted for elution, and the content of the rupatadine fumarate genotoxic impurities in the rupatadine fumarate genotoxic impurities in the rupatadine fumarate genotoxic impurities is detected. Effective separation between rupatadine fumarate and genotoxic impurities and between genotoxic impurities can be achieved, accurate quantitative analysis can be achieved, and the quality of drugs can be effectively controlled.

The invention belongs to the technical field of pharmaceutical analysis, and discloses a method for determining two potential genotoxic impurities in carbon [13C]-urea by gas chromatography. Comprising the following steps: (1) preparing a sample solution; (2) preparing a reference solution; (3) taking nitrogen as carrier gas, and detecting by adopting a medium-polarity or weak-polarity chromatographic column separation system; and (4) calculating by a peak area through an external standard method to obtain the accurate contents of methyl carbamate and ethyl carbamate in the carbon [13C]-urea. The method disclosed by the invention is simple to operate, good in specificity, linearity, precision and accuracy and high in sensitivity, and can realize accurate detection of residual potential genotoxic impurities, namely methyl carbamate and ethyl carbamate, in the carbon [13C]-urea.

The invention relates to the technical field of pharmaceutical analysis, and particularly discloses a high performance liquid chromatography-mass spectrometry tandem method (HPLC-MS / MS) for detecting a genotoxic impurity L-alanine isopropyl ester in an emtricitabine-propofol tenofovir tablet. By optimizing chromatographic column selection, a mobile phase gradient elution procedure and mass spectrometric detection parameters, the method realizes exclusive, sensitive and accurate detection of the L-isopropyl alanine. The methodological verification result shows that the detection limit is 1.00 ng / mL, the quantification limit is 2.00 ng / mL, the linear range is 2.00-100.00 ng / mL (the correlation coefficient r is equal to 0.9994), the recovery rate is stabilized between 92% and 98%, and the precision and repeatability are good. The detection method has the advantages of high sensitivity and strong selectivity, is suitable for quality control of emtricitabine propofol tenofovir tablets, meets the control requirements of ICH M7 guide on genotoxic impurities, and has good practicability and popularization value.

In the field of gene therapy, a major hurdle is the design and identification of constructs and gene therapy vectors providing therapeutic effects while displaying satisfactory safety profiles. In the treatment of Diamond-Blackfan Anemia (DBA), therapies alleviating several crucial anemia symptoms, such as blood or bone marrow cellularity, hemoglobin levels, erythrocytes levels, or platelet levels, while showing satisfactory safety profiles remain a challenge. The present invention provides constructs encoding ribosomal protein genes involved in DBA, such as genes encoding RPS19, RPS17, RPS24, RPS10, RPL35a, RPL11, RPS26, and RPL5, vectors, methods, cells, and medical uses thereof, addressing these challenges and finding particular applications in the field of autologous cell therapy treatment of DBA. Further, the present invention provides a non-genotoxic conditioning protocol for preparing a subject prior to cell therapy treatment for DBA using construct of the present invention.

This invention relates to the field of pharmaceutical impurity preparation technology, and discloses a method for synthesizing apixaban genotoxic impurity I. The steps include: S1. performing an addition reaction of compound II in an alkaline system to generate compound III; S2. subjecting the product of S1 to elimination and isomerization reactions under heating, separating the reaction product to obtain apixaban genotoxic impurity I. The synthesis method of this invention has high safety, low raw material cost, and can achieve a one-pot reaction while ensuring yield and product purity (no product separation and purification is required between each reaction step; only one separation and purification is needed after obtaining the final product).

Described herein is an adeno-associated virus (AAV) complex platform including an asymmetrically modified inverted terminal repeat (ITR). The AAV complex has advantages of increased productivity and expression efficiency of a transgene, and decreased genotoxicity, by having an asymmetric ITR in which any one of two ITRs is modified. Also, described herein is a composition comprising the adeno-associated virus complex and a method of gene therapy.

The invention provides a method for detecting the genetic toxicity of nitrosamine compounds, and belongs to the technical field of biological detection. According to the method, key conditions of a detection system are systematically optimized, so that the detection sensitivity and the result reliability of the genotoxicity of the nitrosamine compounds are remarkably improved, and the mutagenicity which is difficult to detect under conventional conditions can be stably and clearly identified; the establishment of the method provides powerful technical support for early screening and safety risk assessment of nitrosamine impurities in drugs and chemicals, and has important practical application value for guaranteeing medication safety and product quality.

This invention discloses a method for detecting genotoxic impurities in fosfomycin calcium. The method involves preparing a test solution and a reference solution using hydrochloric acid as a solvent, and then detecting the test solution and reference solution using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS) to obtain the content of genotoxic impurities in fosfomycin calcium. The genotoxic impurities are: 1,2-epoxypropyl diethyl phosphate, (2-propenyl)-diethyl phosphate, allyl diethyl phosphate, and propyl diethyl phosphate. This invention develops an analytical method capable of simultaneously detecting four diethyl phosphonate genotoxic impurities in fosfomycin calcium. This method exhibits good specificity, injection precision, linear range, limit of quantitation, limit of detection, accuracy, repeatability, intermediate precision, and robustness. It can be used for the detection and monitoring of diethyl phosphonate impurities in fosfomycin calcium raw materials, ensuring product quality and improving the safety of clinical medication.

The invention belongs to the technical field of analysis and detection, and discloses a method for detecting impurities in a tadalafil bulk drug and / or a tadalafil intermediate. The method for detecting the impurities in the tadalafil bulk drug and / or the tadalafil intermediate comprises the following steps: taking the tadalafil bulk drug and / or the tadalafil intermediate to be detected, and preparing a sample solution; taking an impurity standard substance, and preparing a standard sample solution; and detecting the impurities in the sample solution by adopting an ultra-high performance liquid chromatography-mass spectrometry method. According to the detection method, two N-nitroso genotoxic impurities and ten other process impurities in tadalafil raw materials and / or tadalafil intermediates can be determined at the same time, the specificity is high, the sensitivity is high, results of multiple different types of impurities can be obtained at the same time in one-time determination, the analysis efficiency is effectively improved, and the method is suitable for popularization and application. Powerful guarantee is provided for analyzing generation factors influencing impurities, effectively controlling medicine quality and guaranteeing medicine use safety.

The invention discloses a benzimidazole derivative as well as a preparation method and application thereof. The benzimidazole derivative has a thiophene-benzimidazole structure and is a compound shown in a structural formula I. In the structural formula I, n is an integer from 0 to 3, R1 is C1-6 alkyl, and R2 is selected from at least one of hydrogen, C1-6 alkyl, C1-6 alkenyl, C1-6 alkynyl, C1-6 alkoxy, C1-6 alkylamino, C1-4 halogenated alkyl and a substituted or unsubstituted pyrazole ring. According to the benzimidazole derivative disclosed by the invention, through a thiophene-benzimidazole structure, the benzimidazole derivative has larger Stokes shift; in addition, no obvious genotoxicity is shown on cells, and gene mutation induction is reduced; the method can be better used for DNA dyeing.

The invention discloses a photodegradable impurity in a betamethasone sodium phosphate injection as well as a preparation method and a detection method of the photodegradable impurity. The structure of the photodegradation impurity I in the betamethasone sodium phosphate injection is as shown in the following formula I. The impurity has a potential genotoxic structure, can be generated in a large quantity under the illumination condition, can stably exist in a medicine, and seriously affects the quality of the betamethasone sodium phosphate injection. According to the invention, the structure is clarified, so that a clear control standard can be formulated for the impurity, and the impurity can be conveniently monitored and the quality of the betamethasone sodium phosphate injection can be conveniently controlled.

Here, the inventors developed a strategy to modify the chromatin status of HSPCs at the HBG promoters to reactivate HbF expression. In particular, the inventors used from 1 to 4 single guide RNA (sgRNA) molecules spanning from the -220 to the -20 region of the HBG promoters together with CRISPR-based epigenome editors, namely dCas9-CBPcore and Tet1-dCas9. The first editor is capable of inserting histone acetylation while the second editor is performing DNA demethylation. Both epigenetic marks are associated with active transcription and are present in fetal erythroid cells expressing HbF. This strategy allows the ex vivo modification of adult HSCs to recreate a fetal-like epigenetic context leading to HbF production. Moreover, it avoids the genotoxicity associated with classical genome editing tools relying on DNA cleavage. Thus, the present invention relates to a method of increasing the expression of gamma globin in a population of eukaryotic cells through epigenome editing.

The invention relates to a detection method for toxic impurities of clomacobatel and application, and belongs to the technical field of medicine quality control. The technical problem to be solved is to provide a method for simultaneously separating and detecting four genotoxic impurities, namely p-methyl benzyl chloride, p-(chloromethyl) benzyl alcohol, p-chloromethyl benzaldehyde and p-chloromethyl benzoic acid, from a clomacobamate raw material medicine. The key points of the technical scheme are as follows: chromatographic conditions are as follows: a chromatographic column is a chromatographic column of ion exchange and reverse phase silica gel mixed filler; a mobile phase A is a phosphate buffer solution; a mobile phase B is acetonitrile; the elution conditions are as follows: when the elution time is 0 min, the volume ratio of the mobile phase A to the mobile phase B is 4: 1, 0-30 min, the volume ratio of the mobile phase A to the mobile phase B is changed to 1: 4, 30-35 min, and the volume ratio of the mobile phase A to the mobile phase B is maintained to be 1: 4. The method is good in system applicability, low in detection limit and good in repeatability.

Acute exposure to estrogenic chemicals is shown herein to induce DNA damage mediated by formation of ERα-dependent R-loops. Disclosed herein are methods for evaluating the safety and activity of a xenoestrogen. Also disclosed are methods for identifying subject particularly susceptible to the genotoxic effects of endogenous estrogens or environmental xenoestrogens. This method can be used to select suitable hormone therapies, such as a selective estrogen receptor modulator (SERM), aromatase inhibitor, or selective estrogen receptor degrader (SERD).

The invention discloses an HPLC (High Performance Liquid Chromatography) method for detecting the content of two genotoxic impurities in phentolamine mesylate, belonging to the technical field of pharmaceutical analysis. The method comprises the following steps: 1) preparing a test solution and a reference solution; 2) setting high performance liquid detection conditions: adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filler, taking an ammonium acetate buffer solution as a mobile phase A, taking acetonitrile as a mobile phase B, and carrying out gradient elution; 3) respectively and precisely sucking the test solution and the reference solution, injecting into a liquid chromatograph, and recording a chromatogram chart.According to the method, the two genotoxic impurities in the phentolamine mesylate can be rapidly, effectively, accurately and reliably separated and detected, the product quality of the phentolamine mesylate is improved, and the medication safety of a patient is further improved.

The invention discloses a method for detecting the content of three potential genotoxic impurities in metoprolol tartrate. The method comprises the following steps: detecting a sample to be detected by adopting gas chromatography-mass spectrometry (GC-MS); the parameter conditions of the gas chromatography are as follows: the temperature of a sample inlet is 210-230 DEG C; the heating procedure is as follows: the initial temperature is 35-45 DEG C, and the temperature is kept for 4-6 minutes; the temperature is raised to 235-245 DEG C at the temperature raising speed of 10-20 DEG C / min, and the temperature is kept for 1.5-2.5 min; headspace parameters are as follows: the temperature of a heating box is 85-95 DEG C; the temperature of a quantitative loop is 120-130 DEG C; the temperature of the transmission line is 125-135 DEG C; the balance time of the headspace bottle is 8-12 min; the sample introduction time is 0.3 to 0.8 min; and the GC circulation time is 25 to 30 minutes. The method disclosed by the invention is simple to operate and high in detection speed, and has the advantages of high sensitivity, high precision and high repeatability.

The invention relates to the technical field of cefotaxime acid impurity control, and particularly discloses a method for controlling genotoxic impurities in cefotaxime acid. Carrying out water quenching and organic solvent extraction on reaction liquid obtained after the reaction of the 7-ACA and the AE active ester, and adjusting the pH value of the obtained water-phase feed liquid to be less than or equal to 7; and then adding a sulfur-loaded adsorbent for adsorption, carrying out solid-liquid separation, and crystallizing the obtained filtrate under an acidic condition to obtain the cefotaxime acid. According to the method provided by the invention, the existing cefotaxime acid synthesis step and crystallization step do not need to be specially set, and the content of 2-mercaptobenzothiazole in the cefotaxime acid product can be ensured to be below 3.0 ppm only by ensuring that the feed liquid before adsorption is an acidic aqueous phase solution and the content of 2-mercaptobenzothiazole in the feed liquid is below 0.5%. The method can be suitable for various cefotaxime acid synthesis processes with a water phase extraction step, is wide in application range, and can realize industrial production.

The present application discloses a benzimidazole derivative, a preparation method therefor, and a use thereof. The benzimidazole derivative of the present application comprises a thiophene-benzimidazole structure, and is a compound represented by structural formula I, where n is an integer of 0-3, R1 is a C1-6 alkyl group, and R2 is selected from at least one of hydrogen, a C1-6 alkyl group, a C1-6 alkenyl group, a C1-6 alkynyl group, a C1-6 alkoxy group, a C1-6 alkylamino group, a C1-4 haloalkyl group, and a substituted or unsubstituted pyrazole ring. The benzimidazole derivative of the present application has a larger Stokes shift thanks to the thiophene-benzimidazole structure. Moreover, the benzimidazole derivative of the present application shows no obvious genotoxicity to cells, which reduces induction of gene mutation, allowing it to be better used for DNA staining.

The invention belongs to the technical field of medicine synthesis, and particularly relates to a preparation method of a high-purity melitracen intermediate 1 and control of genotoxic impurities of the high-purity melitracen intermediate 1. The invention provides a genotoxic impurity A or B, a preparation method thereof and a preparation method of a melitracen intermediate 1. The method has the advantages of high yield, high purity, simple operation and mild conditions, and is suitable for industrial mass production.

The present application relates to the technical field of medicine, in particular to a potential genotoxic impurity in a morpholine nitroimidazole raw material and preparation and a preparation method and application thereof.The potential genotoxic impurity is 2-methyl-5-nitro-1-nitrosoimidazole.A new structure compound 2-methyl-5-nitro-1-nitrosoimidazole of diazoles is found in the synthesis process of the morpholine nitroimidazole raw material for the first time.The compound is identified as a potential genotoxic impurity because of containing a nitrosamine warning structure.A method for analyzing the potential genotoxic impurity 2-methyl-5-nitro-1-nitrosoimidazole by using ultra-high performance liquid chromatography-mass spectrometry is established for the first time, which is of great significance to the quality control of the morpholine nitroimidazole raw material and preparation.The detection method provides a reference for the quality evaluation of other starting materials, intermediates and raw materials that can produce 2-methyl-5-nitro-1-nitrosoimidazole.

Methods, compositions and kits are disclosed for using fluorescence light microscopy methods such as directional genomic hybridization to detect, analyze, and / or measure chromosomal stability and genotoxicity.

An adeno-associated virus (AAV) complex platform is disclosed that includes an asymmetrically modified terminal inverted repeat (ITR). The AAV complex has an asymmetric ITR in which any one of two ITRs is modified, and thus has the advantages of increased productivity and transgene expression efficiency, and reduced genetic toxicity. Also disclosed are compositions comprising the adeno-associated virus complexes and gene therapy methods.

The disclosure compositions and methods for non-genotoxic conditioning with an anti-CD45 antibody, anti-CD45 antibody drug conjugate, or anti-CD45 chimeric antigen receptor expressing T cell. The methods involve altering a CD45 polynucleotide sequence in an HSPC using a base editor to encode a CD45 polypeptide with reduced binding to the antibody. Embodiments of the disclosure also feature compositions and methods for treating diseases and conditions by non-genotoxic conditioning with an anti-CD45 antibody, anti-CD45 antibody drug conjugate, or anti-CD45 chimeric antigen receptor expressing T cell (CAR-T) prior to, simultaneously with, or after transplant of the based edited HSPC. The compositions and methods disclosed herein advantageously can reduce or eliminate the use of genotoxic agents such as busulfan, and provide edited HSPCs that are resistant to the disclosed anti-CD45 antibody, anti-CD45 antibody drug conjugate, and anti-CD45 chimeric antigen receptor expressing T cell.

The invention discloses a sitagliptin phosphate pharmaceutical composition as well as a preparation, a preparation method and application thereof. The pharmaceutical composition comprises sitagliptin phosphate, HP-beta-CD and mesoporous silica. According to the invention, a synergistic effect is generated by utilizing a molecular-level clathration effect of HP-beta-CD and a nano-level confinement adsorption effect of mesoporous silica, so that chemical degradation of sitagliptin phosphate, especially generation of a genotoxic impurity NTTP, is remarkably inhibited from molecular and physical double levels, and the stability of a product is greatly improved. Meanwhile, the mesoporous silica effectively solves the technical problems that HP-beta-CD is strong in hygroscopicity and not easy to directly tablet, and the content uniformity of low-dose drugs is fundamentally ensured by forming uniform composite particles. The process is simple, granulation is not needed, the tablet is suitable for industrial production, the prepared tablet is excellent in stability and high in safety, and animal experiments show that the bioavailability of the tablet is superior to that of a commercially available reference preparation.

Compositions and methods for non-genotoxic monoclonal antibody (mAb) conditioning, where the methods involve altering a cluster of differentiation 117 (CD117; c-KIT) polynucleotide sequence in a hematopoietic stem cell (HSC) or progenitor thereof to encode a CD117 polypeptide with reduced binding to the antibody. In various embodiments, the methods further include introducing a therapeutic alteration to a gene of the HSC or progenitor thereof for treatment of a hemoglobinopathy.

The invention discloses a group of biomarkers for evaluating genotoxicity. The biomarker disclosed by the invention is an RFC5 gene, a CHK1 gene, an RAD9 gene, an RAD53 gene, an RAD6 gene, an RAD5 gene, an RAD30 gene, an RAD52 gene, an XRS2 gene, an RAD50 gene, a PHR1 gene, an RAD1 gene, an NTG2 gene, an RAD27 gene and / or an MSH2 gene. The biomarker disclosed by the invention can be used for evaluating genotoxicity in a sensitive and high-throughput manner by covering a plurality of toxicity pathways, the accuracy rate of the biomarker applied to genotoxicity evaluation in eight negative and positive standard substances reaches 100%, and the biomarker has a very good application prospect.

To develop novel therapeutic techniques that do not pose the risk of genotoxicity to help infertility patients. [Solution] A single-stranded negative-strand RNA viral vector containing a nucleic acid sequence encoding a protein associated with the formation or development of germ cells, or a pharmaceutical composition for preventing or treating infertility containing an RNA molecule encoding the protein, is provided.

The invention relates to a method for determining the level and type of mutation events associated with the use of a targeted genetic modification, such as in the use of designer nucleases, to modify a target site of nucleic acid in a cell or virus population. The method comprises carrying out a mutation event determination on a targeted nucleic acid in a population of modified nucleic acids that have been treated with the targeted genetic modification, and a reference control analysis on a non-targeted nucleic acid. The invention further relates to the use of the method for screening of potential targeted genetic modification agents for therapeutic use and to estimate the genomic integrity and stability of a nucleic acid such as a viral vector or genomic DNA.

The application discloses a method for determining the content of potential genotoxic impurity tert-butyl hydroperoxide in deoxycholic acid bulk drug or an intermediate thereof, which adopts pre-column derivatization high performance liquid chromatography for detection, and adopts triphenylphosphine for derivatization treatment of the sample in pre-column derivatization; the detection conditions of the high performance liquid chromatography are as follows: a chromatographic column is an Agilent Eclipse Plus C18 column or a Waters Symmetry C18 column; a mobile phase A is 0.01% to 0.10% phosphoric acid solution; a mobile phase B is acetonitrile; a flow rate is 0.9 to 1.1 ml / min; a detection wavelength is 210 to 250 nm; a sample injection volume is 5 to 10 ul; a column temperature is 30 to 40 DEG C; a diluent is methanol or acetonitrile; and an elution mode is gradient elution. The method has the characteristics of low cost, simple operation, high accuracy and good sensitivity, and is suitable for quality control of medicines.