Check patentability & draft patents in minutes with Patsnap Eureka AI!

Bispecific antibody constructs for CDH3 and CD3

A bispecific antibody, CDR-H3 technology, applied in the direction of antibodies, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, specific peptides, etc., can solve the problems of inability to exert cytotoxicity and limitations

Active Publication Date: 2018-03-02
AMGEN RES (MUNICH) GMBH
View PDF164 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chimpanzees are considered an endangered species, and because of their human-like nature, the use of this animal for drug safety testing has been banned in Europe and severely restricted elsewhere
[0022] Although T-cell-engaged bispecific single-chain antibodies described in the art have great therapeutic potential for treating malignant diseases, most of these bispecific molecules have the limitation that they are species-specific and only recognize human Antigen, and possible primate i.e. macaque counterpart
Furthermore, the majority of such bispecific molecules are further limited in that they cannot cross species boundaries to fulfill their desired functions, such that they can recognize human and primate homologues but cannot exert e.g. T cell-mediated cytotoxicity

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Bispecific antibody constructs for CDH3 and CD3
  • Bispecific antibody constructs for CDH3 and CD3
  • Bispecific antibody constructs for CDH3 and CD3

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0440] Generation of CHO cells expressing wild-type and chimeric CDH3

[0441] For the construction of chimeric molecules for epitope mapping, the sequences of the corresponding five extracellular domains Doml to Dom5 (or D1 to D5) and their subdomains (A, B, and C) of human CDH3 were replaced by the corresponding mouse domains. sequence replacement. The following 20 molecules were produced; see also figure 1 and 2.

[0442] · Hu CDH3 / Doml mu (aa 108-215) SEQ ID NO: 13

[0443] o Hu CDH3 / DomlA mu(aa 108-143) SEQ ID NO: 14

[0444] o Hu CDH3 / DomlB mu(aa 144-179) SEQ ID NO: 15

[0445] o Hu CDH3 / DomlC mu(aa 180-215) SEQ ID NO: 16

[0446] · Hu CDH3 / Dom2mu (aa 216-327) SEQ ID NO: 17

[0447] o Hu CDH3 / Dom2A mu(aa 216-252) SEQ ID NO: 18

[0448] o Hu CDH3 / Dom2B mu(aa 253-290) SEQ ID NO: 19

[0449] o Hu CDH3 / Dom2C mu(aa 291-327) SEQ ID NO: 20

[0450] · Hu CDH3 / Dom3mu (aa 328-440) SEQ ID NO: 21

[0451] o Hu CDH3 / Dom3A mu(aa 328-363) SEQ ID NO: 22

[0452] o Hu CDH3 / Dom...

Embodiment 2

[0467] Epitope clustering of murine scFv-fragments

[0468] Cells transfected with human or murine CDH3 or with chimeric human / mouse CDH3 molecules (see Example 1) were stained with crude, undiluted periplasmic extracts containing scFvs that bind human / cynomolgus CDH3. Mouse monoclonal anti-FLAG-M2 antibody (1 μg / ml; 50 μl in PBS / 2% FCS; Sigma F1804), followed by anti-mouse IgG Fcγ-PE (1:100, 50 μl; Jackson Immunoresearch #115- 116-071) Detection of bound scFv molecules. All antibodies were diluted in PBS with 2% FCS. As a negative control, cells were incubated with PBS / 2% FCS instead of periplasmic extract. Samples were measured by flow cytometry. The results are shown in Figure 4.

[0469] in particular, Figure 4A Binders recognizing the extracellular domain D1 of human CDH3, more precisely the subdomain D1B are shown (FACS signal loss in the corresponding chimeric CDH3 constructs). Note that the binder designated CDH3-6 is the parental binder of CDH3-4. The binder d...

Embodiment 3

[0472] Biacore-based determination of antibody affinity for human and cynomolgus monkey (cynomolgus) CDH3

[0473] CDH3 target binding of antibodies of the invention was determined by Biacore analysis experiments using recombinant CDH3 (human and cyno CDH3, respectively) fusion proteins with human albumin (HALB).

[0474] Specifically, CM5 sensor chips (GE Healthcare) were immobilized with approximately 600-800 RU of the corresponding recombinant antigens using acetate buffer at pH 4.5 according to the manufacturer's instructions. CDH3xCD3 bispecific antibody samples were loaded at five concentrations: 5OnM, 25nM, 12.5nM, 6.25nM and 3.13nM diluted in HBS-EP running buffer (GE Healthcare). The flow rate was 30 μl / min for 3 minutes, and then HBS-EP running buffer was applied again at a flow rate of 30 μl / ml for 8 min to 20 min. Chip regeneration was performed using 10 mM glycine, 10 mM NaCl solution, pH 1.5. Data sets were analyzed using BiaEval software. Usually two independ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Ec50aaaaaaaaaa
Thermal stabilityaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a bispecific antibody construct comprising a first human binding domain which binds to human CDH3 on the surface of a target cell and a second binding domain which binds to human CDS on the surface of a T cell. Moreover, the invention provides a polynucleotide encoding the antibody construct, a vector comprising said polynucleotide and a host cell transformed or transiected with said polynucleotide or vector. Furthermore, the invention provides a process for the production of the antibody construct of the invention, a medical use of said antibody construct anda kit comprising said antibody construct.

Description

technical field [0001] The present invention relates to a bispecific antibody construct comprising a first human binding domain that binds human CDH3 on the surface of a target cell and a second binding domain that binds human CD3 on the surface of a T cell. In addition, the present invention provides a polynucleotide encoding the antibody construct, a vector comprising the polynucleotide, and a host cell transformed or transfected with the polynucleotide or vector. Furthermore, the invention provides a method for producing the antibody construct of the invention, the medical use of said antibody construct and a kit comprising said antibody construct. Background technique [0002] The cadherin superfamily encompasses more than 100 members in humans, including the so-called canonical cadherins P-cadherin, E-cadherin, N-cadherin and R-cadherin, each of which Active in a single group of tissues (Takeichi M. Development, 102: 639-55 (1988), van Roy F., Nature Rev., V14: 121-134...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/46C07K16/28A61K39/395A61P35/00
CPCC07K16/28C07K16/2809C07K2317/622C07K2317/73C07K2317/92C07K2317/31C07K2317/33A61P35/00A61P35/02C07K2317/94C07K2317/565C07K2317/56A61K2039/505C07K2317/732C07K16/3053
Inventor B·韦斯A·L·弗里斯克R·齐尔茨P·库弗T·劳姆D·劳J·安拉尔R·路特布斯L·纳尔沃德C·达尔霍夫C·布鲁麦尔P·霍夫曼
Owner AMGEN RES (MUNICH) GMBH
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com