	US 11,840,567 B2
	Bispecific antibodies with specific binding to CD47 and PD-L1
Kirill Vladimirovich Solovyev, St.Petersburg (RU); Andrei Borisovich Ulitin, Puschino (RU); Timofey Aleksandrovich Nemankin, St. Petersburg (RU); Valery Vladimirovich Solovyev, Pushchino (RU); and Dmitry Valentinovich Morozov, Saint Petersburg (RU)
Appl. No. 16/753,587
Filed by JOINT STOCK COMPANY “BIOCAD”, St.Petersburg (RU)
PCT Filed Oct. 3, 2018, PCT No. PCT/EA2018/050001 § 371(c)(1), (2) Date Apr. 3, 2020, PCT Pub. No. WO2019/068302, PCT Pub. Date Apr. 11, 2019.
Claims priority of application No. 201791961 (EA), filed on Oct. 3, 2017.
Prior Publication US 2020/0270345 A1, Aug. 27, 2020
Int. Cl. C07K 16/28 (2006.01)

CPC C07K 16/2803 (2013.01) [C07K 16/2827 (2013.01); C07K 2317/31 (2013.01); C07K 2317/55 (2013.01); C07K 2317/565 (2013.01); C07K 2317/622 (2013.01); C07K 2317/732 (2013.01); C07K 2317/734 (2013.01); C07K 2317/76 (2013.01); C07K 2317/92 (2013.01)]

13 Claims

1. A bispecific monoclonal antibody or its antigen-binding fragment, which specifically binds to CD47 and PD-L1, comprising a binding site to CD47, and at least one binding site to PD-L1, characterized in that the binding site to CD47 includes one of:

a) (i) a heavy chain variable domain that comprises CDR1, CDR2 and CDR3 sequences, wherein CDR1 comprises SEQ ID NO: 1, wherein CDR2 comprises SEQ ID NO: 6 or 7, wherein CDR3 comprises SEQ ID NO: 17, and

(ii) a light chain variable domain comprising the sequences CDR1, CDR2, CDR3, wherein:

CDR1 comprises SEQ ID NO: 22, CDR2 comprises SEQ ID NO: 36, and CDR3 comprises SEQ. ID NO: 50;

CDR1 comprises SEQ ID NO: 25, CDR2 comprises SEQ ID NO: 38, and CDR3 comprises SEQ. ID NO: 53;

CDR1 comprises SEQ ID NO: 29, CDR2 comprises SEQ ID NO: 48, and CDR3 comprises SEQ. ID NO: 64;

CDR1 comprises SEQ ID NO: 28, CDR2 comprises SEQ ID NO: 40, and CDR3 comprises SEQ. ID NO: 56;

CDR1 comprises SEQ ID NO: 23, CDR2 comprises SEQ ID NO: 37, and CDR3 comprises SEQ. ID NO: 51;

CDR1 comprises SEQ ID NO: 30, CDR2 comprises SEQ ID NO: 42, and CDR3 comprises SEQ. ID NO: 58;

CDR1 comprises SEQ ID NO: 24, CDR2 comprises SEQ ID NO: 43, and CDR3 comprises SEQ. ID NO: 59; or

CDR1 comprises SEQ ID NO: 32, CDR2 comprises SEQ ID NO: 45, and CDR3 comprises SEQ. ID NO: 61;

b) (i) a heavy chain variable domain comprising the sequences CDR1, CDR2, CDR3, where CDR1 comprises SEQ. ID NO: 1, where CDR2 comprises SEQ. ID NO: 8, SEQ. ID NO: 9, SEQ. ID NO: 11, or SEQ. ID NO: 12, where CDR3 comprises SEQ. ID NO: 17; and

(ii) a light chain variable domain comprising the sequences CDR1, CDR2, CDR3, where CDR1 comprises SEQ ID NO: 32, CDR2 comprises SEQ ID NO: 45, and CDR3 comprises SEQ. ID NO: 61; and

c) wherein the binding site to CD47 is an anti-CD47 VHH variable domain, comprising a heavy chain variable domain comprising CDR1, CDR2 and CDR3, wherein CDR1 comprises SEQ ID NO: 2 or SEQ ID NO: 4, CDR2 comprises SEQ. ID NO: 10 or SEQ. ID NO: 15, and CDR3 comprises SEQ. ID NO: 18 or SEQ ID NO: 20; and

the binding site to PD-L1 comprises:

a heavy chain variable domain that comprises CDR1, CDR2, CDR3 sequences, wherein CDR1 comprises SEQ ID NO: 5, wherein CDR2 comprises SEQ ID NO: 16 and wherein CDR3 comprises SEQ ID NO: 21; and

a light chain variable domain that comprises CDR1, CDR2, CDR3 sequences, wherein CDR1 comprises SEQ ID NO: 35, wherein CDR2 comprises SEQ ID NO: 49 and wherein CDR3 comprises SEQ ID NO: 65.