# US 9,518,998 B2 · Drug target capturing method

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- Number: US 9,518,998 B2
- Document ID: US9518998B2
- Status: Granted
- Kind: B2
- Application number: 14/945,438
- Filed: 2015-11-19
- Published: 2016-06-02
- Granted: 2016-12-13
- Earliest claimed: 2013-05-21
- Assignees: Hitgen Inc.
- Applicants: HitGen LTD.
- Inventors: Jin Li; Benyanzi Yang; Dengfeng Dou; Xiaohu Ge; Hongmei Song; Jinqiao Wan; Yan Zhang; Xiao Hu; Xing Wang; Jingchao Feng; Guoqing Zhong
- Web page: https://patentgrep.com/patent/US9518998B2

## CPC

- G01N 33/94: involving narcotics {or drugs or pharmaceuticals, neurotransmitters or associated receptors}
- C12N 15/09: Recombinant DNA-technology
- C12Q 1/6811: Selection methods for production or design of target specific oligonucleotides or binding molecules
- G01N 33/582: {with fluorescent label}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention discloses a method for capturing drug targets, comprising the following steps of: (1) preparing row materials, i.e., compounds and DNA or RNA or one party of other specific affinity materials; (2) linking: covalently linking the compounds to the DNA or RNA or the one party of other specific affinity materials, to obtain labeled compounds; (3) transferring: transferring the labeled compounds obtained in the step (2) into cells by a specific gene transfer method; (4) capturing targets: disrupting the cells in the step (3), and then capturing the DNA or RNA in the step (1) by immobilized complementary DNA or RNA, or capturing the one party of affinity materials in the step (1) by the other party of immobilized specific affinity materials, thus to enrich the targets through the covalently linked labeled compounds and the affinity of the targets; (5) identifying targets: dissociating the enriched targets from a stationary phase, deploying the targets by a gel electrophoresis method or by an equivalent separation method, and comparing differential proteins by proteomics thus to identify the potential targets; and (6) determining targets: expressing or purchasing the targets identified in the step (5) by priority, and comparing by interaction of the compounds with those targets one by one to determine the targets for the compounds. By the method of the present invention, compounds with low membrane permeability may be transferred into cells to be bound with the targets in the cells, to realize the target identification closer to the real physiological environment and thus provide more conclusive evidences for the acting mechanism of the compounds to diseases. Additionally, the method for capturing drug targets of the present invention is novel, easy, efficient, and low in cost, and has good application prospects.

## Claims

### Claim 1

A method for capturing drug targets, comprising the following steps of:
(1) Preparing a raw material comprising drug compounds and DNA, RNA or one part of other specific affinity materials;
(2) Linking: covalently linking the drug compounds to the DNA, RNA or the one part of other specific affinity materials, to obtain labeled compounds;
(3) Transferring: transferring the labeled compounds obtained in the step (2) into cells;
(4) Capturing targets: disrupting the cells in the step (3), and then capturing the DNA or RNA in the step (1) by immobilized complementary DNA or RNA, or capturing the one part of specific affinity materials in the step (1) by the immobilized other part of the specific affinity materials, thus to enrich the targets;
(5) Identifying targets; and
(6) Determining targets.

### Claim 2 (depends on 1)

The method according to claim 1, characterized in that,
the target identification described in the step (5) is to dissociate the enriched targets from a stationary phase, deploy the enriched targets by a gel electrophoresis method, and compare differential proteins by proteomics thus to identify the potential targets; and
the target determination described in the step (6) is to express the targets identified in the step (5) by priority, and compare by interaction of the compounds with those targets one by one to determine the targets for the compounds.

### Claim 3 (depends on 1)

The method according to claim 1, characterized in that, in the step (1), the molecular weight of the compounds ranges from 100 Da to 4000 Da.

### Claim 4 (depends on 1)

The method according to claim 1, characterized in that the DNA or RNA in the step (1) has a functional group for the covalent linkage to the compounds in the step (2).

### Claim 5 (depends on 1)

The method according to claim 1, characterized in that, in the step (1), the length of the DNA or RNA is not less than five base pairs or bases.

### Claim 6 (depends on 1)

The method according to claim 1, characterized in that, in the step (1), the DNA or RNA is tagged with biotin or fluorescein or isotope or other tags for tracing.

### Claim 7 (depends on 1)

The method according to claim 1, characterized in that the one part of other specific affinity materials in the step (1) is one part of any pair of specific affinity materials having covalent linkage to the compounds in the step (2).