Anti-cd93 constructs and uses thereof

Abstract

This application provides an anti-CD93 construct (e.g., an anti-CD93 antibody) that binds to CD93, a nucleic acid molecule encoding the amino acid sequence of the anti-CD93, a vector containing the nucleic acid molecule, a host cell containing the vector, a method for preparing the anti-CD93 construct, a pharmaceutical composition containing the anti-CD93 construct, and a method for using the anti-CD93 construct or the composition.

Description

Technical field

[0001] The present invention relates to anti-CD93 constructs, such as anti-CD93 antibodies, and uses thereof.

Prior Technology

[0002] CD93 (cluster of differentiation 93) is a protein encoded by the CD93 gene in humans. CD93 is a C-type lectin transmembrane receptor that plays a role not only in the process of cell-cell adhesion but also in host defense. CD93 was originally thought to be the receptor of C1q, but it is now thought to be actually related to intercellular adhesion and clearance of apoptotic cells. The intracellular cytoplasmic tail of this protein contains two highly conserved domains that may be related to the function of CD93. Indeed, the highly charged juxtamembrane domain has been found to interact with moesins, proteins known to play a role in linking transmembrane proteins to the cytoskeleton and in remodeling the cytoskeleton. This process appears to be critical for adhesion, migration and phagocytosis.

[0003] The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety.

Content of invention

[0004] The following summary is illustrative only and is not intended to be limiting in any way. That is, the following summary is provided to introduce the most salient aspects, benefits and advantages of the novel molecules and their uses. Accordingly, the following summary is not intended to identify essential features of the claimed subject matter, nor is it intended to be used to determine the scope of the claimed subject matter.

[0005] In one aspect, the application provides an anti-CD93 construct comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with an antibody portion comprising Antibodies or antibody fragments of the second heavy chain variable region (VH-2) and the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein: a) the VH-2 comprises SEQ ID NO: HC-CDR1 of the amino acid sequence of 1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and the VL-2 Including LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 6; b ) The VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 19 HC-CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and the amine comprising SEQ ID NO: 22 LC-CDR3 of amino acid sequence; c) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34 and comprising SEQ ID HC-CDR3 of the amino acid sequence of NO: 35, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 36, and LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 And LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; d) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 49, comprising the amino acid of SEQ ID NO: 50 HC-CDR2 of sequence and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 51, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 52, comprising SEQ ID NO: 53 LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 54 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 54; e) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, comprising HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 67, and the VL-2 comprises LC comprising the amino acid sequence of SEQ ID NO: 68 - CDR1, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 70; f) the VH-2 comprises an amine comprising SEQ ID NO: 81 The HC-CDR1 of the amino acid sequence, the HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and the HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 83, and the VL-2 comprises the amino acid sequence comprising SEQ ID NO LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 84, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 85, and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 86; g) the VH-2 Including HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 97, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 98 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 99, and The VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 100, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 101 and LC comprising the amino acid sequence of SEQ ID NO: 102 - CDR3; h) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 113, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114 and the amine comprising SEQ ID NO: 115 The HC-CDR3 of the amino acid sequence, and the VL-2 comprises the LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 116, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 117 and the LC-CDR2 comprising the amino acid sequence of SEQ ID NO LC-CDR3 of the amino acid sequence of: 118; i) the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 129, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 130 And HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 131, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 132, comprising the amino acid sequence of SEQ ID NO: 133 LC-CDR2 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 134; j) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, comprising SEQ ID NO: 146 HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 147 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the VL-2 comprises LC- comprising the amino acid sequence of SEQ ID NO: 148, 355 or 358 CDR1, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or 356 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 150, 357 or 359; k) the VH-2 comprises comprising SEQ ID NO: HC-CDR1 of the amino acid sequence of 161, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 162 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 163, and the VL-2 Including LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 164, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 165 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 166; l ) The VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178 and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 179 HC-CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 180 or 353, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 181 or 354 and comprising SEQ ID NO LC-CDR3 of the amino acid sequence of: 182; m) the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 193, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 194 And HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 195, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 196, comprising the amino acid sequence of SEQ ID NO: 197 LC-CDR2 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 198; n) the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 209, comprising SEQ ID NO: 210 HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 211 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 211, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 212, comprising SEQ LC-CDR2 comprising the amino acid sequence of ID NO: 213 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 214; or o) the VH-2 comprises a LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 289 HC-CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 291, and the VL-2 comprises the amine comprising SEQ ID NO: 292 LC-CDR1 comprising the amino acid sequence, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293, and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294; p) the VH-2 comprises comprising SEQ ID NO: HC-CDR1 of the amino acid sequence of 17 or 304, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and The VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, 301, 302, 303 or 306, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and comprising SEQ ID NO: 22 amino acid sequences of LC-CDR3.

[0006] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 3, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 4, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 6, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0007] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 or 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and VL comprising i ) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, 301, 302, 303 or 306, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising SEQ ID NO: LC-CDR3 of an amino acid sequence of 22, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0008] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 35, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 36, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0009] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 49, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 50, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 51, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 52, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 53 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 54, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0010] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 67, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 68, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 70, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0011] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 82, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 83, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 84, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 85 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 86, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0012] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 97, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 98, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 99, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 100, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 101 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 102, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0013] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 113, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 115, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 116, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 117 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 118, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0014] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 129, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 130, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 131, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 132, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 133 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 134, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0015] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 146, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 147, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 148, 355 or 358, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or 356 and iii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 150, 357 or 359 LC-CDR3 of the amino acid sequence, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0016] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 161, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 162 and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 163, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 164, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 165 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 166, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0017] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 179, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 180 or 353, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 181 or 354 and iii) LC comprising the amino acid sequence of SEQ ID NO: 182 - CDR3, or a variant thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0018] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 193, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 194, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 195, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 196, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 197 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 198, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0019] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 209, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 210, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 211, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 212, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 213 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 214, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0020] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 291, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 292, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0021] In some embodiments, the VH comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 or 304, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305, and comprising the amino acid sequence of SEQ ID NO: 18 or 305 The HC-CDR3 of the amino acid sequence of NO: 19, or its variant comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and VL comprises SEQ ID NO: 20, LC-CDR1 of the amino acid sequence of 301, 302, 303 or 306, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0022] In another aspect, the present application comprises an anti-CD93 construct comprising an antibody portion specifically binding to CD93, the antibody portion comprising: a) respectively comprising HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the VH chain region of the sequence, and HC-CDR1, HC-CDR2, and HC-CDR3 in the VL chain region comprising the sequence shown in SEQ ID NO: 14, respectively. LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3; b) comprising a VH chain having a sequence shown in any one of SEQ ID NO: 29 and 307-312, respectively HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the region, and HC-CDR1, HC-CDR2, and HC-CDR3 comprising the sequence shown in any one of SEQ ID NO: 30 and 313-318, respectively. LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region; c) respectively comprising CDR1 in the VH chain region having the sequence shown in SEQ ID NO: 45 HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of , CDR2, and CDR3, and amine groups comprising CDR1, CDR2, and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 46, respectively LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence; d) HC-CDR1 comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 61, respectively , HC-CDR2 and HC-CDR3, and LC-CDR1, LC-CDR2 and LC-CDR1, LC-CDR2 and LC-CDR1 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 62 CDR3; e) HC-CDR1, HC-CDR2, and HC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2, and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 77, and respectively comprising LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region of the sequence shown in SEQ ID NO: 78; HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 within the VH chain region of the sequence shown, and VL chains having the sequence shown in SEQ ID NO: 94, respectively LC-CDR1, LC-CDR2, and LC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the region; g) CDR1, CDR2 in the VH chain region comprising the sequence shown in SEQ ID NO: 109, respectively and HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequence of CDR3, and the amino acid sequences of CDR1, CDR2, and CDR3 respectively comprising the VL chain region having the sequence shown in SEQ ID NO: 110 LC-CDR1, LC-CDR2 and LC-CDR3; h) HC-CDR1, HC comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 125, respectively - CDR2 and HC-CDR3, and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 126; i) HC-CDR1, HC-CDR2 and HC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 141, and respectively comprising LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region of the sequence shown in NO: 142; j) comprise having SEQ ID NO: 157 and 360- HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the VH chain region of the sequence shown in any one of 362, and respectively comprising SEQ ID NO: 158 and LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region of the sequence shown in any one of 363-365; HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the VH chain region of the sequence shown in : 173, and comprising the sequence shown in SEQ ID NO: 174, respectively LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region; HC-CDR1, HC-CDR2 and HC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region of the sequence shown, and include any one of SEQ ID NO: 190 and 350-352, respectively LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region of the sequence shown in the sequence; m) respectively comprising the sequence shown in SEQ ID NO: 205 HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequences of CDR1, CDR2, and CDR3 in the VH chain region, and CDR1, HC-CDR1, and CDR1 in the VL chain region having the sequence shown in SEQ ID NO: 206, respectively LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequences of CDR2 and CDR3; n) respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 221 HC-CDR1, HC-CDR2, and HC-CDR3 of the acid sequence, and LC-CDR1, LC-CDR1, LC-CDR2 and LC-CDR3; o) HCs comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in any one of SEQ ID NO: 287 and 319-321, respectively - CDR1, HC-CDR2 and HC-CDR3, and respectively comprising the amino acid sequence of CDR1, CDR2 and CDR3 within the VL chain region having the sequence shown in any one of SEQ ID NO: 288 and 322-324 LC-CDR1, LC-CDR2 and LC-CDR3; p) respectively comprising the amino acids of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in any one of SEQ ID NO: 307-312 HC-CDR1, HC-CDR2, and HC-CDR3 of the sequence, and amino acids comprising CDR1, CDR2, and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 313-318, respectively LC-CDR1, LC-CDR2, and LC-CDR3 of the sequence; or q) comprise the amines of CDR1, CDR2, and CDR3, respectively, within the VH chain region having the sequence shown in any one of SEQ ID NO: 319-321 HC-CDR1, HC-CDR2, and HC-CDR3 of the amino acid sequence, and respectively comprise the amines of CDR1, CDR2, and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 322-324 LC-CDR1, LC-CDR2 and LC-CDR3 of amino acid sequence.

[0023] In some embodiments according to any one of the above anti-CD93 constructs, wherein the VH comprises SEQ ID NO: 13, 29, 45, 61, 77, 93, 109, 125, 141, 157, 173, 189 , 205, 221, 287, 307-312 and any one of the amino acid sequence of 319-321, or comprising at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96% , 97%, 98% or 99% in any one of the amino acid sequence variants of sequence identity; and / or wherein the VL comprises SEQ ID NO: 14, 30, 46, 62, 78, 94, The amino acid sequence of any one of 110, 126, 142, 158, 174, 190, 206, 222, 288, 313-318 and 322-324 or comprising at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 13, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 14, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 29 and 307-312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%). , 96%, 97%, 98% or 99% in any one of the amino acid sequence variants of sequence identity; and VL comprises the amine group of any one of SEQ ID NO: 30 and 313-318 Acid sequence, or comprising amino acids having at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity variants of the sequence. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 45, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 46, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 61, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or any one of 99%) sequence identity variants of amino acid sequences; and VL comprises the amino acid sequence of SEQ ID NO: 62, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 77, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 78, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 93, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 94, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 109, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or any one of 99%) sequence identity variants of amino acid sequences; and VL comprises the amino acid sequence of SEQ ID NO: 110, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 125, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 126, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 141, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 142, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 157, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 158, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 173, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 174, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 189 and 347-349, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%. , 96%, 97%, 98% or 99% in any one of the amino acid sequence variant of sequence identity; and VL comprises the amine group of any one of SEQ ID NO: 190 and 350-352 Acid sequence, or comprising amino acids having at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity variants of the sequence. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 205, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 206, or comprises a variant having at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 221, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 222, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 287 and 319-321, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%. , 96%, 97%, 98% or 99% in any one of) the variant of the amino acid sequence of sequence identity; And VL comprises the amine group of any one in SEQ ID NO: 288 and 322-324 Acid sequence, or comprising amino acids having at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity variants of the sequence.

[0024] In some embodiments according to any one of the above anti-CD93 constructs, the antibody moiety is an antibody or antigen-binding fragment thereof selected from the group consisting of full length antibodies, bispecific antibodies, single chain Fv (scFv) fragment, Fab fragment, Fab' fragment, F(ab')2, Fv fragment, disulfide bond stabilized Fv fragment (dsFv), (dsFv)2, Fv-Fc fusion, scFv-Fc fusion , scFv-Fv fusion, bifunctional antibody, trifunctional antibody and tetrafunctional antibody. In some embodiments, the antibody portion is a full length antibody.

[0025] In some embodiments according to any of the above anti-CD93 constructs, the antibody portion has an Fc fragment selected from the group consisting of IgG, IgA, IgD, IgE, IgM, and combinations and hybrids thereof The Fc fragment of the mixture. In some embodiments, the Fc fragment is selected from the group consisting of Fc fragments from IgGl, IgG2, IgG3, IgG4, and combinations and hybrids thereof. In some embodiments, the Fc fragment has reduced effector function compared to a corresponding wild-type Fc fragment. In some embodiments, the Fc fragment has enhanced effector functions compared to a corresponding wild-type Fc fragment. In some embodiments, the Fc fragment has an extended serum half-life. In some embodiments, the Fc fragment has a reduced serum half-life.

[0026] In some embodiments according to any of the above anti-CD93 constructs, the antibody partially blocks the binding of CD93 to IGFBP7, such as human IGFBP7.

[0027] In some embodiments according to any of the anti-CD93 constructs above, the antibody partially blocks CD93 binding to MMRN2, such as human MMRN2.

[0028] In some embodiments according to any of the above anti-CD93 constructs, the antibody partially blocks a) CD93 binding to IGFBP7 and / or b) CD93 binding to MMRN2.

[0029] In some embodiments according to any of the anti-CD93 constructs above, the CD93 is human CD93.

[0030] In some embodiments, a fusion protein comprising any of the anti-CD93 constructs described above is provided. In some embodiments, an anti-CD93 construct is fused to one or more cell signaling peptides or proteins. In some embodiments, an anti-CD93 construct is fused to one or more VEGF binding moieties. In some embodiments, the anti-CD93 construct is fused to one or more VEGF-A binding moieties. In some embodiments, the VEGF-A binding moiety is Aflibercept. In one embodiment, the fusion protein comprises a heavy chain fusion polypeptide comprising the amino acid sequence shown in SEQ ID NO: 366, or comprising at least about 80% (such as at least about 80%, 85%) , 90%, 95%, 96%, 97%, 98% or 99% in any one of) the variant of the amino acid sequence of sequence identity; And light chain polypeptide, this light chain polypeptide comprises SEQ ID NO: The amino acid sequence shown in 367, or comprising any one having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) A variant of an amino acid sequence of sequence identity.

[0031] In another aspect, the present application provides a pharmaceutical composition comprising any one of the above-mentioned anti-CD93 constructs and a pharmaceutically acceptable carrier.

[0032] In another aspect the application provides an isolated nucleic acid encoding any of the anti-CD93 constructs described above.

[0033] In another aspect the application provides a vector comprising any of the above isolated nucleic acids.

[0034] In another aspect the application provides an isolated host cell comprising any of the above isolated nucleic acids or vectors.

[0035] In another aspect the application provides an immunoconjugate comprising any of the above anti-CD93 constructs linked to a therapeutic agent or label.

[0036] In another aspect, the present application provides a method for producing an anti-CD93 construct, which comprises: a) cultivating the isolated host as in technical scheme 25 under the condition of effectively expressing the anti-CD93 construct cells; and b) obtaining the expressed anti-CD93 construct from the host cell.

[0037] In another aspect, the present application provides a method of treating a disease or condition in an individual, comprising administering to the individual an effective amount of any of the above-mentioned anti-CD93 constructs or pharmaceutical compositions. In some embodiments, the disease or condition is associated with abnormal vascular structure. In some embodiments, the disease or condition is cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer comprises CD93+ endothelial cells. In some embodiments, the cancer comprises IGFBP7+ blood vessels. In some embodiments, the cancer is characterized by tumor hypoxia. In some embodiments, the cancer is locally advanced or metastatic cancer. In some embodiments, the cancer is selected from the group consisting of lymphoma, colorectal cancer, brain cancer, breast cancer, ovarian cancer, endometrial cancer, esophageal cancer, prostate cancer, cervical cancer, kidney cancer, bladder cancer, Gastric cancer, non-small cell lung cancer, melanoma and pancreatic cancer. In some embodiments, an anti-CD93 construct is administered to a subject parenterally. In some embodiments, the method further comprises administering a second therapy. In some embodiments, the second therapy is selected from the group consisting of surgery, radiation, gene therapy, immunotherapy, bone marrow transplant, stem cell transplant, hormone therapy, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy and chemotherapy. In some embodiments, the second therapy is immunotherapy. In some embodiments, immunotherapy comprises administering an immunomodulator. In some embodiments, the immune modulator is an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor comprises an anti-PD-L1 antibody or an anti-PD-1 antibody. In some embodiments, the individual is human.

Implementation

[0093] CROSS-REFERENCE TO RELATED APPLICATIONS

[0094] This application claims U.S. Provisional Application 63 / 058,359 filed on July 29, 2020, U.S. Provisional Application 63 / 084,474 filed on September 28, 2020, and National Application No. 63 / 084,474 filed on June 2, 2021. Priority to PCT / US2021 / 035542 and National Application No. PCT / US2021 / 043784 filed July 29, 2021, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0095] The present application provides novel anti-CD93 constructs (such as anti-CD93 monoclonal or multispecific antibodies) that specifically bind to CD93, methods of preparing such anti-CD93 constructs, methods of using such constructs (such as method of treating a disease or condition).

[0096] Anti-CD93 antibodies (such as anti-CD93 antibodies that block the interaction between CD93 and IGFBP7) can effectively treat tumors or cancer, block abnormal tumor angiogenesis, normalize immature and leaky tumor blood vessels, and promote tumor growth. function of the vascular network, promotes vascular maturation, promotes a favorable tumor microenvironment, increases immune cell infiltration in tumors, increases tumor perfusion, reduces proliferation in tumors, sensitizes tumors to second therapies, and / or facilitates second agents of delivery. See eg WO2021062128A1, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the anti-CD93 constructs described herein reduce tumor size. In some embodiments, the anti-CD93 constructs described herein promote immune cell infiltration in tumors. In some embodiments, the anti-CD93 constructs described herein promote vascular maturation in tumors. In some embodiments, the anti-CD93 constructs described herein sensitize a tumor to a second therapy or facilitate delivery of a second agent. I. Definition

[0097] The term "antibody" is used in its broadest sense and encompasses various antibody structures including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (such as bispecific antibodies), full-length antibodies, and An antigen-binding fragment thereof is sufficient as long as it exhibits the desired antigen-binding activity. The term "antibody portion" refers to a full-length antibody or an antigen-binding fragment thereof.

[0098] A full-length antibody comprises two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable domains of the heavy and light chains can be referred to as "VH" and "VL", respectively. The variable regions in both chains generally contain three highly variable loops called complementarity determining regions (CDRs) (including light chain (LC) CDRs of LC-CDR1, LC-CDR2 and LC-CDR3, including HC- Heavy chain (HC) CDR of CDR1, HC-CDR2 and HC-CDR3). The CDR boundaries of the antibodies and antigen-binding fragments disclosed herein can be defined or identified according to the Kabat, Chothia or Al-Lazikani conventions (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three CDRs of the heavy or light chain are inserted between flanking stretches called the framework regions (FRs), which are more conserved than the CDRs and form a scaffold that supports the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding, but exhibit various effector functions. Antibodies are classified based on the amino acid sequence of the constant region of their heavy chains. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG and IgM, characterized by the presence of alpha, delta, epsilon, gamma and mu heavy chains, respectively. Several major antibody classes are divided into subclasses such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), lgA1 (α1 heavy chain) or lgA2 (α2 heavy chain) .

[0099] As used herein, the term "antigen-binding fragment" refers to antibody fragments including, for example, diabody, Fab, Fab', F(ab')2, Fv fragments, disulfide bond-stabilized Fv fragments ( dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide bond-stabilized diabodies (ds diabodies), single chain Fv (scFv), scFv dimers (divalent functional antibody), a multispecific antibody formed from a portion of an antibody comprising one or more CDRs, a camelid single domain antibody, a nanobody, a domain antibody, a bivalent domain antibody, or any antibody that binds to an antigen but does not comprise a complete antibody structure Other antibody fragments. Antigen-binding fragments are capable of binding to the same antigen as the parent antibody or parent antibody fragment (eg, parent scFv). In some embodiments, an antigen-binding fragment may comprise one or more CDRs from a particular human antibody grafted to a framework region from one or more different human antibodies.

[0100] "Fv" is the minimal antibody fragment that contains a complete antigen recognition and antigen binding site. This fragment consists of a dimer of one heavy chain variable region domain and one light chain variable region domain in tight, non-covalent association. These two domains fold to result in six hypervariable loops (3 loops each for the heavy and light chains) that contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only the three CDRs specific for an antigen) is capable of recognizing and binding antigen, but usually with a lower affinity than the entire binding site.

[0101] "Single-chain Fv" (also abbreviated "sFv" or "scFv") is an antibody fragment comprising the VH and VL antibody domains linked into a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the structure required for antigen binding. For a review of scFv, see Plückthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0102] As used herein, the term "CDR" or "complementarity determining region" is intended to mean the non-contiguous antigen combining sites found within the variable regions of heavy and light chain polypeptides. Such specific regions have been described in: Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987); Al-Lazikani B. et al., J. Mol. Biol., 273: 927-948 (1997); MacCallum et al. , J. Mol. Biol. 262:732-745 (1996); Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Lefranc M.P. et al., Dev. Comp. Immunol., 27: 55- 77 (2003); and Honegger and Plückthun, J. Mol. Biol., 309:657-670 (2001), wherein the definitions include overlapping amino acid residues or subsets of amino acid residues when compared to each other. However, application of either definition to refer to an antibody or a CDR of a grafted antibody or a variant thereof is intended to be within the scope of the term as defined and used herein. The amino acid residues encompassing the CDRs as defined by each of the references cited above are set forth in Table 1 below for comparison. CDR prediction algorithms and interfaces are known in the art, including, for example, Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Ehrenmann F. et al., Nucleic Acids Res., 38: D301- D307 (2010); and Adolf-Bryfogle J. et al., Nucleic Acids Res., 43: D432-D438 (2015). The contents of the references cited in this paragraph are incorporated herein by reference in their entirety for use in this application and may be included in one or more technical solutions herein. In some embodiments, the CDR sequences provided herein are based on IMGT definitions. For example, CDR sequences can be obtained by the VBASE2 tool (http: / / www.vbase2.org / vbase2.php, see also Retter I, Althaus HH, Münch R, Müller W: VBASE2, an integrative V gene database. Nucleic Acids Res. 2005 Jan 1; 33 (database album): D671-4, which is hereby incorporated by reference in its entirety) assay. Table 1: CDR Definitions Kabat 1 Chothia 2 MacCallum 3 IMGT 4 AHo 5 V H CDR1 31-35 26-32 30-35 27-38 25-40 V H CDR2 50-65 53-55 47-58 56-65 58-77 V H CDR3 95-102 96-101 93-101 105-117 109-137 V L CDR1 24-34 26-32 30-36 27-38 25-40 V L CDR2 50-56 50-52 46-55 56-65 58-77 V L CDR3 89-97 91-96 89-96 105-117 109-137 1 Residue numbering follows the above-mentioned nomenclature of Kabat et al. 2 Residue numbering follows the above-mentioned nomenclature of Chothia et al. 3 Residue numbering follows the above-mentioned nomenclature of MacCallum et al. 4 Residue numbering follows the above-mentioned nomenclature of Lefranc et al. 5 Residues Base numbers follow the nomenclature of Honegger and Plückthun above

[0103] The expression "numbering of variable domain residues as in Kabat" or "numbering of amino acid positions as in Kabat" and its variants refer to the heavy chain variable domains or Numbering system for light chain variable domains. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortenings of, or insertions into, FRs or hypervariable regions (HVRs) of the variable domains. For example, a heavy chain variable domain may include a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and an inserted residue after heavy chain FR residue 82 (e.g., according to Kabat , residues 82a, 82b and 82c, etc.). For a given antibody, the Kabat numbering of residues can be determined by aligning the homologous regions of the antibody sequence to the "standard" Kabat numbering sequence.

[0104] Unless otherwise indicated herein, the numbering of residues in immunoglobulin heavy chains is as in the EU index in Kabat et al., supra. "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody.

[0105] "Framework" or "FR" residues are variable domain residues other than CDR residues as defined herein.

[0106] "Humanized" forms of non-human (eg, rodent) antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. Humanized antibodies are, for the most part, human immunoglobulins (recipient antibodies) having the desired antibody specificity, affinity and capacity in which residues from the hypervariable regions (HVRs) of the recipient are modified from a non-human species. Substitution of residues in hypervariable regions (such as mouse, rat, rabbit or non-human primate) of the (donor antibody). In some instances, framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. In addition, humanized antibodies may comprise residues which are not found in either the recipient antibody or the donor antibody. These modifications are made to further refine antibody potency. In general, a humanized antibody will comprise substantially all of at least one, and usually two, variable domains, in which all or substantially all hypervariable loops correspond to those of a non-human immunoglobulin and in which all or substantially all FR is the FR of human immunoglobulin sequence. A humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically, that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992) .

[0107] A "human antibody" is an antibody whose amino acid sequence corresponds to that of an antibody produced by a human and / or has been prepared using any of the techniques for preparing human antibodies as disclosed herein. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues. Human antibodies can be generated using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). The methods described in: Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1): 86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be produced by administering antigens to transgenic animals that have been modified to produce such antibodies in response to antigenic challenge but whose endogenous loci have been disabled, e.g., immunized Xenomices (see, eg, US Patent Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology). See also, eg, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies produced via human B cell fusion tumor technology.

[0108] "Amino acid sequence identity (%)" or "homology" with respect to the polypeptide and antibody sequences identified herein is defined as a sequence where any conservative substitutions are considered part of the sequence identity. After alignment, the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the compared polypeptides. Alignment for the purpose of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR) or MUSCLE software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. However, for the purposes herein, the sequence comparison computer program MUSCLE was used to generate % amino acid sequence identity values ​​(Edgar, R.C., Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1) :113, 2004).

[0109] "Homologous" refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. When one position in both of the two compared sequences is occupied by the same base or amino acid monomer subunit, for example if one position in each of the two protein molecules is occupied by lysine or if both If one position in each of the DNA molecules is occupied by adenine, the molecules are homologous at that position. The % homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of compared positions times 100. For example, two sequences are 60% homologous if 6 / 10 positions in the two sequences match or are homologous. For example, the protein sequence SGTSTD (SEQ ID NO: 421 ) shares 50% identity with TGTSDA (SEQ ID NO: 422). In general, comparisons are made when two sequences are aligned for maximum homology.

[0110] The term "constant domain" refers to that portion of an immunoglobulin molecule that has a more conserved amino acid sequence than the rest of the immunoglobulin (ie, the variable domain) that contains the antigen combining site. The constant domain comprises the CH1, CH2, and CH3 domains (collectively CH) of the heavy chain and the CHL (or CL) domain of the light chain.

[0111] The "light chains" of antibodies (immunoglobulins) from any mammalian species can be assigned to two groups called kappa ("κ") and lambda ("λ") based on the amino acid sequences of their constant domains. One of two distinct types.

[0112] A "CH1 domain" (also known as "C1" of an "H1" domain) generally extends from about amino acid 118 to about amino acid 215 (EU numbering system).

[0113] "Hinge region" is generally defined as the region in IgG extending from Glu216 to Pro230 corresponding to human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). Hinge regions of other IgG isotypes can be aligned to the IgGl sequence by placing the first and last cysteine ​​residues that form the S-S bonds between the heavy chains at the same positions.

[0114] The "CH2 domain" (also referred to as the "C2" domain) of the human IgG Fc region generally extends from about amino acid 231 to about amino acid 340. The CH2 domain is unique in that it is not closely paired with another domain. Instead, two N-linked branched carbohydrate chains are inserted between the two CH2 domains of the intact native IgG molecule. It has been hypothesized that carbohydrates may provide a surrogate for domain-domain pairing and help stabilize the CH2 domain. Burton, Molec Immunol. 22:161-206 (1985).

[0115] A "CH3 domain" (also referred to as a "C2" domain) comprises a stretch of the C-terminal residues to the CH2 domain in the Fc region (i.e., from about amino acid residue 341 of the antibody sequence to the C-terminus, The C-terminus is usually located at amino acid residue 446 or 447 of IgG).

[0116] The terms "Fc region" or "fragment crystallizable region" are used herein to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. While the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the human IgG heavy chain Fc region is generally defined as extending from an amino acid residue at position Cys226 or from Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, eg, during antibody production or purification, or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, compositions of intact antibodies may comprise antibody populations from which all K447 residues have been removed, antibody populations from which no K447 residue has been removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Suitable native sequence Fc regions for the antibodies described herein include human IgGl, IgG2 (IgG2A, IgG2B), IgG3 and IgG4.

[0117] "Fc receptor" or "FcR" describes a receptor that binds the Fc region of an antibody. Preferred FcRs are native sequence human FcRs. In addition, preferred FcRs are receptors that bind IgG antibodies (gamma receptors) and include receptors of the FcγRI, FcγRII, FcRN, and FcγRIII subclasses, including allele variants and alternatively spliced ​​forms of these receptors, FcγRII receptors These include FcyRIIA ("activating receptor") and FcyRIIB ("inhibiting receptor"), which have similar amino acid sequences and differ mainly in their cytoplasmic domains. Activating receptor FcyRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain. (See M. Daëron, Annu. Rev. Immunol. 15:203-234 (1997). FcRN is critical for the recycling of antibodies to the blood, allowing for prolonged serum half-life of antibodies. FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol.9: 457-92 (1991); Capel et al., Immunomethods 4: 25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126: 330-41 (1995). Other FcRs, including FcRs identified in the future, are encompassed herein by the term "FcR".

[0118] As used herein, the term "epitope" refers to a specific atom or group of amino acids on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, they are likely to bind the same epitope within the antigen.

[0119] As used herein, a first antibody or fragment thereof inhibits target antigen binding of a second antibody or fragment thereof by at least about 50% (such as at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%), the first antibody or its fragment and the second antibody or its fragment "competes" for binding to the target antigen, or vice versa. A high-throughput method based on cross-competition antibody "binning" is described in PCT Publication No. WO 03 / 48731.

[0120] As used herein, the term "specifically binds", "specifically recognizes" or "has specificity for" refers to a measurable and reproducible interaction (such as between a target and an antibody or antibody portion). binding), which determines the presence or absence of a target in the presence of a heterogeneous population of molecules, including biomolecules. For example, an antibody or antibody portion that specifically recognizes a target (which may be an epitope) is an antibody or antibody portion that binds this target with greater affinity, avidity, ease and / or duration than it binds to other targets. In some embodiments, the antibody binds to an unrelated target to an extent that is about 10% less than the antibody binds to that target, as measured, eg, by radioimmunoassay (RIA). In some embodiments, the dissociation constant (KD) of the antibody that specifically binds to the target is ≤10-5M, ≤10-6M, ≤10-7M, ≤10-8M, ≤10-9M, ≤10-10M, ≤ 10-11M or ≤ 10-12M. In some embodiments, the antibody specifically binds to an epitope on a protein that is conserved among proteins from different species. In some embodiments, specific binding can include exclusive binding, but need not be. The binding specificity of an antibody or antigen binding domain can be determined experimentally by methods known in the art. Such methods include, but are not limited to, Western blotting, ELISA-, BLI, RIA-, ECL-, IRMA-, EIA-, BIACORE™ tests, and peptide scanning.

[0121] As used herein, a molecule A (eg, an anti-CD93 construct as described herein) "blocking" the binding of a molecule B (eg, CD93) to a molecule C (eg, IGFBP7 or MMRN2) refers to both direct and indirect blocking. broken. For example, instead of directly blocking the binding of CD93 to IGFBP7 or MMRN2 by occupying at least a portion of the binding site on CD93 responsible for binding IGFBP7 or MMRN2, anti-CD93 constructs as described herein can be obtained by altering the structure of CD93 such that The inability of CD93 to bind to IGFBP7 / MMRN2 blocks the binding of CD93 to IGFBP7 or MMRN2.

[0122] An "isolated" or "purified" antibody (or construct) is an antibody (or construct) that has been identified, separated and / or recovered (eg, native or recombinant) from a component of the environment in which it was produced. Preferably, an isolated polypeptide is free from all other components from the environment in which it was produced.

[0123] An "isolated" nucleic acid molecule encoding a construct, antibody, or antigen-binding fragment thereof described herein is one that has been identified and separated from at least one contaminating nucleic acid molecule with which it is normally associated in the environment in which it is produced. Preferably, an isolated nucleic acid is free from all components related to the environment in which it was produced. Isolated nucleic acid molecules that encode the polypeptides and antibodies described herein are in a form other than that found in nature. Isolated nucleic acid molecules therefore are distinguished from nucleic acid encoding the polypeptides and antibodies described herein that occur naturally in cells. Isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location other than its natural chromosomal location.

[0124] The term "control sequences" refers to DNA sequences required for the expression of an operably linked coding sequence in a particular host organism. Control sequences suitable for prokaryotes include, for example, a promoter, an optional operator sequence, and a ribosome binding site. Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.

[0125] A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, the DNA of a presequence or secretory leader sequence is operably linked to the DNA of a polypeptide if it appears to be a preprotein involved in the secretion of the polypeptide; if a promoter or enhancer affects the transcription of a coding sequence, it is operably linked to that sequence. operably linked; or, if a ribosomal binding site is positioned so as to facilitate translation, it is operably linked to the coding sequence. Generally, "operably linked" means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading frame. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers are used according to conventional practice.

[0126] As used herein, the term "vector" refers to a nucleic acid molecule capable of transmitting another nucleic acid to which it has been linked. The term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which they have been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vehicles are referred to herein as "expression vehicles."

[0127] As used herein, the term "transfection" or "transformation" or "transduction" refers to a method of transferring or introducing exogenous nucleic acid into a host cell. A "transfected" or "transformed" or "transduced" cell is a cell that has been transfected, transformed or transduced with an exogenous nucleic acid. Cells include primary individual cells and their progeny.

[0128] The terms "host cell", "host cell strain" and "host cell culture" are used interchangeably and refer to a cell into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells may include "transformants" and "transformed cells", which include primary transformed cells and progeny derived therefrom, regardless of the number of passages. The nucleic acid content of the progeny may not be identical to that of the parental cells and may contain mutations. Included herein are mutant progeny that have the same function or biological activity as screened or selected for on the original transformed cell.

[0129] The term "immunoconjugate" includes reference to the covalent linkage of a therapeutic agent or detectable label to an antibody, such as an antibody moiety described herein. Linkage can be direct or indirect via a linker, such as a peptide linker.

[0130] As used herein, "treatment" or "treating" is a method used to obtain beneficial or desired results, including clinical results. For purposes of this application, a beneficial or desired clinical outcome may include, but is not limited to, one or more of the following: alleviation of one or more symptoms caused by the disease, reduction of the extent of the disease, stabilization of the disease (e.g., arrest or delay) exacerbation of the disease), prevent or delay the spread of the disease (such as metastasis), prevent or delay the recurrence of the disease, delay or slow the progression of the disease, improve the disease state, provide remission (partial or total) of the disease, reduce one or more treatments Doses of other drugs needed for the disease, to delay the progression of the disease, to enhance or improve quality of life, to increase weight gain and / or to prolong survival. "Treatment" also encompasses the reduction of pathological consequences of cancer, such as tumor volume. The methods of the present application encompass any one or more of these treatment aspects.

[0131] In the context of cancer, the term "treatment" includes any or all of the following: inhibition of cancer cell growth, inhibition of cancer cell replication, reduction of overall tumor burden, and amelioration of one or more symptoms associated with the disease.

[0132] The term "inhibition" or "inhibit" refers to the reduction or cessation of any phenotypic characteristic, or the reduction or cessation of the incidence, degree or likelihood of that characteristic. "Reducing" or "inhibiting" means reducing, reducing or arresting activity, function and / or amount compared to a reference. In certain embodiments, "reduce" or "inhibit" means capable of causing an overall reduction of 20% or greater. In another embodiment, "reduce" or "inhibit" means capable of causing an overall reduction of 50% or greater. In another embodiment, "reduce" or "inhibit" means capable of causing an overall reduction of 75%, 85%, 90%, 95% or greater.

[0133] As used herein, a "reference" refers to any sample, standard or level used for comparison purposes. References can be obtained from healthy and / or disease-free samples. In some embodiments, a reference can be obtained from an unprocessed sample. In some instances, a reference is obtained from a disease-free or untreated sample of an individual. In some embodiments, a reference is obtained from one or more healthy individuals who are not the individual or patient.

[0134] As used herein, "delaying disease progression" means delaying, hindering, slowing, stifling, stabilizing, arresting and / or delaying the progression of a disease such as cancer. This delay can be of varying lengths, depending on the disease being treated and / or the individual's medical history. As will be apparent to those skilled in the art, a sufficient or substantial delay may actually encompass prophylaxis such that the individual does not develop the disease. For example, advanced cancer, such as metastatic development, can be delayed.

[0135] As used herein, "prevention" includes providing prophylaxis against the onset or recurrence of a disease in an individual who may be susceptible to the disease but has not yet been diagnosed with the disease.

[0136] As used herein, to "inhibit" a function or activity is to reduce the function or activity when compared to an otherwise identical condition except for the relevant condition or parameter or compared to another condition. For example, an antibody that inhibits tumor growth reduces the rate of tumor growth compared to the rate of tumor growth in the absence of the antibody.

[0137] The terms "subject", "individual" and "patient" are used interchangeably herein to refer to mammals, including but not limited to humans, bovines, horses, felines Animals, canines, rodents or primates. In some embodiments, the individual is human.

[0138] The "effective amount" of a medicament refers to the amount effective to achieve the desired therapeutic or prophylactic result at the desired dose and time period. The specific dosage will vary depending on one or more of the particular compound chosen, the dosing regimen followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to be imaged, and the physical delivery system for its delivery.

[0139] The "therapeutically effective amount" of the substance / molecule, agonist or antagonist of the present application may vary according to factors such as: the individual's disease state, age, sex and weight, and the substance / molecule, agonist The ability of an agent or antagonist to elicit a desired response in an individual. A therapeutically effective amount is also one such that any toxic or detrimental effects of the substance / molecule, agonist or antagonist, are outweighed by the therapeutically beneficial effects. A therapeutically effective amount can be delivered in one or more administrations.

[0140] "Prophylactically effective dose" refers to the amount that can effectively achieve the desired preventive effect at the necessary dose and time period. Usually, but not necessarily, the prophylactically effective amount will be less than the therapeutically effective amount due to the prophylactic dose being used by the individual before or at an early stage of the disease.

[0141] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to a preparation that is in a form that permits the biological activity of the active ingredient to be effective and that is free of other components that are unacceptably toxic to the subject to whom the formulation is administered. point. Such formulations can be sterile.

[0142] "Pharmaceutically acceptable carrier" means a non-toxic solid, semi-solid or liquid filling known in the art used with a therapeutic agent together to constitute a "pharmaceutical composition" for administration to a subject. agent, diluent, encapsulating material, formulation aid or carrier. Pharmaceutically acceptable carriers are nontoxic to recipients at the dosages and concentrations employed and are compatible with the other ingredients of the formulation. Pharmaceutically acceptable carriers are suitable for the formulation used.

[0143] A "sterile" formulation is sterile or substantially free of viable microorganisms and their spores.

[0144] Administration in "combination" with one or more other therapeutic agents includes simultaneous (concurrent) administration as well as sequential or sequential administration in any order.

[0145] The term "concurrently" is used herein to refer to the administration of two or more therapeutic agents, wherein at least a portion of the administrations overlap in time or wherein one therapeutic agent is administered relative to the other. within a short period of time invested. For example, two or more therapeutic agents are administered no more than about 60 minutes apart, such as no more than about any of the following: 30, 15, 10, 5, or 1 minute.

[0146] The term "sequentially" is used herein to refer to the administration of two or more therapeutic agents in which the administration of one or more agents continues after an interruption of the administration of one or more other agents. For example, the administration of two or more therapeutic agents is over about 15 minutes, such as about 20 minutes, 30 minutes, 40 minutes, 50 minutes, or 60 minutes, 1 day, 2 days, 3 days, 1 week, Administration at intervals of 2 weeks, or 1 month, or longer.

[0147] As used herein, "in combination" refers to the administration of one treatment modality in addition to another treatment modality. Thus, "in combination" refers to the administration of one treatment modality to a subject before, during or after administration of another treatment modality.

[0148] As used herein, the term "package insert" is used to refer to instructions commonly included in commercial packages of therapeutic products, which contain information about the indications, usage, dosage, administration, combination, Information on treatments, contraindications and / or warnings.

[0149] An "article of manufacture" is any manufacture (e.g., a packaging or container) or set. In certain embodiments, an article of manufacture or a kit is marketed, distributed, or sold as a unit for performing the methods described herein.

[0150] It is to be understood that embodiments of the application described herein include "consisting of" and / or "consisting essentially of" the embodiments.

[0151] As used herein, reference to "about" a value or parameter includes (and describes) variations with respect to that value or parameter itself. For example, description referring to "about X" includes description of "X".

[0152] As used herein, reference to "not being" a value or parameter generally means and describes "other than" a value or parameter. For example, a method not for treating type X cancer means that the method is for treating a type of cancer other than type X.

[0153] The term "about X to Y" as used herein has the same meaning as "about X to about Y".

[0154] As used herein and in the appended claims, the singular forms "a" or "an" and "the" include plural referents unless the context clearly dictates otherwise. II. Anti-CD93 Constructs

[0155] The present application provides anti-CD93 constructs comprising an anti-CD93 antibody portion that specifically binds to CD93 as described herein.

[0156] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 1 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and the VL-2 comprises the amino acid comprising SEQ ID NO: 4 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 5, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 6.

[0157] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 3, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 4, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 6, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 7, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 8 and iii) comprising SEQ ID NO : HC-CDR3 of the amino acid sequence of 9, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 10 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 11 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 12, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0158] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 and iii) comprising the amino acid sequence of SEQ ID NO: 3 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 6.

[0159] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 14.

[0160] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 13, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0161] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 17 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the VL-2 comprises the amino acid comprising SEQ ID NO: 20 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 21, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 22.

[0162] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 19, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 20, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 23, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 24, and iii) comprising the amino acid sequence of SEQ ID NO: 24 and iii) comprising SEQ ID NO HC-CDR3 of an amino acid sequence of : 25, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 26 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 27 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 28, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0163] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 and iii) comprising the amino acid sequence of SEQ ID NO: 19 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 22.

[0164] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 29 and 307-312, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 30 and 313-318 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0165] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 29 and 307-312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%). , 95%, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of SEQ ID NO: 30 and 313-318 or comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity Variants of amino acid sequences.

[0166] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 33 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the VL-2 comprises the amino acid comprising SEQ ID NO: 36 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 37, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 38.

[0167] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 35, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 36, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 39, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 40 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :41, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 42 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 43 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 44, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0168] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34 and iii) comprising the amino acid sequence of SEQ ID NO: 35 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 36, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 38.

[0169] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 46.

[0170] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 45, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0171] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 49 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 50 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 51, and the VL-2 comprises the amino acid comprising SEQ ID NO: 52 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 53, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 54.

[0172] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 49, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 50, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 51, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 52, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 53 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 54, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 55, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 56, and iii) comprising the amino acid sequence of SEQ ID NO: 56 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :57, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 58 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 59 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 60, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0173] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 49, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 50 and iii) comprising the amino acid sequence of SEQ ID NO: 51 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 52, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 53 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 54.

[0174] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 62.

[0175] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 61, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0176] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 65 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 67, and the VL-2 comprises the amino acid comprising SEQ ID NO: 68 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 69, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 70.

[0177] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 67, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 68, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 70, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 71, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 72, and iii) comprising the amino acid sequence of SEQ ID NO: 72 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :73, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 74 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 75 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 76, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0178] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and iii) comprising the amino acid sequence of SEQ ID NO: 67 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 68, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 70.

[0179] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 78.

[0180] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 77, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0181] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 81 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 83, and the VL-2 comprises the amino acid comprising SEQ ID NO: 84 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 85, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 86.

[0182] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 82, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 83, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 84, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 85 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 86, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 87, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 88 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :89, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 90 The LC-CDR1 of the amino acid sequence of, ii) comprise the LC-CDR2 of the amino acid sequence of SEQ ID NO:91 and iii) comprise the LC-CDR3 of the amino acid sequence of SEQ ID NO:92, or its in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0183] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 81, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 82 and iii) comprising the amino acid sequence of SEQ ID NO: 83 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 84, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 85 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 86.

[0184] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 94.

[0185] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 93, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0186] In some embodiments, an anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 97 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 98 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 99, and the VL-2 comprises the amino acid comprising SEQ ID NO: 100 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 101, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 102.

[0187] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 97, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 98, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 99, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 100, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 101 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 102, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 103, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 104, and iii) comprising the amino acid sequence of SEQ ID NO: 104 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 105, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 106 The LC-CDR1 of the amino acid sequence of, ii) comprise the LC-CDR2 of the amino acid sequence of SEQ ID NO: 107 and iii) comprise the LC-CDR3 of the amino acid sequence of SEQ ID NO: 108, or its in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0188] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 97, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 98 and iii) comprising the amino acid sequence of SEQ ID NO: 99 HC-CDR3, and VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 100, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 101 and iii) comprising SEQ ID NO : LC-CDR3 of the amino acid sequence of 102.

[0189] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 110.

[0190] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 109, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0191] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of the second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 113 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 115, and the VL-2 comprises the amino acid comprising SEQ ID NO: 116 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 117, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 118.

[0192] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 113, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 115, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 116, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 117 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 118, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 119, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 120 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 121, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 122 The amino acid sequence of LC-CDR1, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 123 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 124, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0193] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 113, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114 and iii) comprising the amino acid sequence of SEQ ID NO: 115 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 116, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 117 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 118.

[0194] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 126.

[0195] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 125, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0196] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 129 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 130 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 131, and the VL-2 comprises the amino acid comprising SEQ ID NO: 132 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 133, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 134.

[0197] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 129, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 130, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 131, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 132, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 133 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 134, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 135, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 136 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 137, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 138 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 139 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 140, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0198] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 129, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 130 and iii) comprising the amino acid sequence of SEQ ID NO: 131 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 132, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 133 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 134.

[0199] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 142.

[0200] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 141, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0201] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 145 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 146 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 147, and the VL-2 comprises SEQ ID NO: 148, 355 or 358 LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 149, LC-CDR2 or 356 comprising the amino acid sequence of SEQ ID NO: 149, and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 150, 357 or 359.

[0202] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 146, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 147, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 148, 355 or 358, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or 356 and iii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 150, 357 or 359 LC-CDR3 of the amino acid sequence, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 151, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 152 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 153, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 154 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 155 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 156, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0203] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 145, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 146 and iii) comprising the amino acid sequence of SEQ ID NO: 147 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 148, 355 or 358, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or 356 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 150, 357 or 359.

[0204] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 157 and 360-362, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 158 and 363-365 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0205] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 157 and 360-362, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%). , 95%, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of SEQ ID NO: 158 and 363-365 or comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity Variants of amino acid sequences.

[0206] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 157, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0207] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 360, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0208] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 360, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0209] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 360, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0210] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 361, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0211] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 361, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0212] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 361, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0213] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 362, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0214] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 362, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0215] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 362, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0216] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 161 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 162 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 163, and the VL-2 comprises the amino acid comprising SEQ ID NO: 164 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 165, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 166.

[0217] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 161, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 162, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 163, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 164, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 165 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 166, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 167, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 168, and iii) comprising the amino acid sequence of SEQ ID NO: 168 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 169, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 170 The LC-CDR1 of the amino acid sequence of, ii) comprise the LC-CDR2 of the amino acid sequence of SEQ ID NO: 171 and iii) comprise the LC-CDR3 of the amino acid sequence of SEQ ID NO: 172, or its in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0218] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 161, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 162 and iii) comprising the amino acid sequence of SEQ ID NO: 163 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 164, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 165 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 166.

[0219] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 174.

[0220] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 173, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0221] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 177 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 179, and the VL-2 comprises the amino acid comprising SEQ ID NO: 180 LC-CDR1 or 353 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 181 or 354, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 182.

[0222] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 179, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 180 or 353, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 181 or 354 and iii) LC comprising the amino acid sequence of SEQ ID NO: 182 - CDR3, or a variant thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of: 179, and VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 180, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 181 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 182. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 183, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 184 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 185, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 186 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 187 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 188, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0223] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 177, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 178 and iii) comprising the amino acid sequence of SEQ ID NO: 179 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 180 or 353, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 181 or 354, and iii ) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 182.

[0224] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 189 and 347-349, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 190 and 350-352 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0225] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 189 and 347-349, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%). , 95%, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of SEQ ID NO: 190 and 350-352 or comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity Variants of amino acid sequences.

[0226] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 189, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0227] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 347, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0228] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 347, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0229] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 347, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% in any of the variants of the amino acid sequence of sequence identity; and VL comprises the amino acid sequence of any one in SEQ ID NO: 352, or comprises A variant of an amino acid sequence having at least about any of 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity.

[0230] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 348, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0231] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 348, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0232] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 348, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0233] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 349, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0234] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 349, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0235] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 349, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0236] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 193 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 194 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 195, and the VL-2 comprises the amino acid comprising SEQ ID NO: 196 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 197, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 198.

[0237] In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 193, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 194, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 195, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 196, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 197 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 198, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 199, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 200 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 201, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 202 The LC-CDR1 of the amino acid sequence of, ii) comprise the LC-CDR2 of the amino acid sequence of SEQ ID NO: 203 and iii) comprise the LC-CDR3 of the amino acid sequence of SEQ ID NO: 204, or its in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0238] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 193, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 194 and iii) comprising the amino acid sequence of SEQ ID NO: 195 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 196, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 197 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 198.

[0239] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 206.

[0240] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 205, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0241] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 includes HC- comprising the amino acid sequence of SEQ ID NO: 209 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 210 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 211, and the VL-2 comprises the amino acid comprising SEQ ID NO: 212 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 213, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 214.

[0242] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 209, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 210, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 211, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 212, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 213 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 214, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 215, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 216, and iii) comprising the amino acid sequence of SEQ ID NO: 216 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :217, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 218 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 219 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 220, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0243] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 209, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 210 and iii) comprising the amino acid sequence of SEQ ID NO: 211 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 212, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 213 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 214.

[0244] In some embodiments, the antibody portion comprises HC-CDR1, HC-CDR2, and HC-CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 222.

[0245] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 221, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0246] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC- comprising the amino acid sequence of SEQ ID NO: 289 CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 291, and the VL-2 comprises the amino acid comprising SEQ ID NO: 292 LC-CDR1 of the sequence, LC-CDR2 of the amino acid sequence of SEQ ID NO: 293, and LC-CDR3 of the amino acid sequence of SEQ ID NO: 294.

[0247] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290, and iii) comprising HC-CDR3 of the amino acid sequence of SEQ ID NO: 291, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID LC-CDR1 of the amino acid sequence of NO: 292, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294, or It is a variant comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 295, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 296 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :297, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 298 The LC-CDR1 of the amino acid sequence of, ii) comprise the LC-CDR2 of the amino acid sequence of SEQ ID NO: 299 and iii) comprise the LC-CDR3 of the amino acid sequence of SEQ ID NO: 300, or its in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0248] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and iii) comprising the amino acid sequence of SEQ ID NO: 291 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 294.

[0249] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 287 and 319-321, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 288 and 322-324 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0250] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 287 and 319-321, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%). , 95%, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of SEQ ID NO: 288 and 322-324 or comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity Variants of amino acid sequences.

[0251] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 287 and 319-321, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 288 and 322-324 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0252] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NOs: 319-321, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%). %, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of the amino acids in SEQ ID NO: 322-324 sequence, or an amino acid sequence comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity of variants.

[0253] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 319, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0254] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 319, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0255] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 319, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0256] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 320, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0257] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 320, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0258] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 320, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0259] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 321, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0260] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 321, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0261] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 321, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0262] In some embodiments, the anti-CD93 construct comprises an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region ( VH-2) and an antibody or antibody fragment of a second light chain variable region (VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises an amino acid sequence comprising SEQ ID NO: 17 or 304 HC-CDR1, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the VL-2 comprising SEQ ID NO: 20 , LC-CDR1 of the amino acid sequence of 301, 302, 303 or 306, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0263] In some embodiments, the VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 or 304, ii) an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i ) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, 301, 302, 303 or 306, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising SEQ ID NO: LC-CDR3 of an amino acid sequence of 22, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0264] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0265] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0266] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 302, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0267] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 303, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0268] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 306, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0269] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0270] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 301, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0271] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 302, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0272] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 303, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0273] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 305, and iii ) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the anti-CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 306, ii) comprising the amine of SEQ ID NO: 21 The amino acid sequence of LC-CDR2 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22.

[0274] In some embodiments, the antibody portion comprises an amino acid sequence comprising CDR1, CDR2 and CDR3 within the VH chain region having the sequence set forth in any one of SEQ ID NO: 29 and 307-312, respectively HC-CDR1, HC-CDR2 and HC-CDR3; and respectively comprising the amine groups of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in any one of SEQ ID NO: 30 and 313-318 LC-CDR1, LC-CDR2 and LC-CDR3 of the acid sequence.

[0275] In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NOs: 307-312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%). %, 96%, 97%, 98% or 99%) sequence identity variants of amino acid sequences; and VL comprises any one of the amino acids in SEQ ID NO: 313-318 sequence, or an amino acid sequence comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity of variants.

[0276] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0277] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0278] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0279] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0280] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0281] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 307, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0282] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0283] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0284] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0285] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0286] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0287] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 308, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0288] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0289] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0290] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0291] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0292] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0293] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 309, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0294] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0295] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0296] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0297] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0298] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0299] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 310, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0300] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0301] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0302] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0303] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0304] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0305] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 311, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0306] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0307] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0308] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0309] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0310] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%). , 98% or 99% in any one of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 312, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%, 96%, 97%. , 98% or 99% in any of the variants of the amino acid sequence of sequence identity; %, 85%, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0312] In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) HC- comprising the amino acid sequence of SEQ ID NO: 33 CDR1; ii) HC-CDR2 comprising the amino acid sequence of RIFPGDGDX1X2YX3GKFKG (SEQ ID NO: 233), wherein X1X2 is AN or TD, and / or X3 is N or D; and iii) comprises TGAAYX1FDPFPY (SEQ ID NO: 234 ), wherein X1 is D or E; and the VL comprises i) LC-CDR1 comprising the amino acid sequence SSX1KSLLHSX2GX3TYLY (SEQ ID NO: 235), wherein X1 is S or T, X2 Be N or S, and / or X3 is V or I; ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37; and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38 .

[0313] In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) a protein comprising the amino acid sequence X1YWX2N (SEQ ID NO: 236) HC-CDR1, wherein X1 is S or T, and / or X2 is L or M; ii) HC-CDR2 comprising the amino acid sequence of RIX1PGDGDX2X3YX4GKFKG (SEQ ID NO: 237), wherein X1 is Y or F, and X2X3 is TD or AN, and / or X4 is N or D; and iii) HC-CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 35, 163 and 179; and the VL comprises i) comprising X1X2X3KSLLHSX4GX5TYLY ( LC-CDR1 of the amino acid sequence of SEQ ID NO: 238), wherein X1X2X3 is SSS, SST or RFS, X4 is N or S, and / or X5=V or I; ii) comprises the amino acid sequence X1MSNLAS (SEQ ID NO: 239) LC-CDR2, wherein X1 is R or Q; and iii) LC-CDR3 comprising the amino acid sequence AQX1LEX2PX3T (SEQ ID NO: 240), wherein X1 is M or N, X2 is R or L , and / or X3 is F or W. In some embodiments, the LC-CDR3 comprises an amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 38, 166, and 182.

[0314] In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) a protein comprising the amino acid sequence X1YVX2H (SEQ ID NO: 241) HC-CDR1, wherein X1 is A or S, and / or X2 is M or I; ii) HC-CDR2 comprising the amino acid sequence of YIX1PYX2DX3TX4YNEKFKG (SEQ ID NO: 242), wherein X1 is F or N, and X2 is N or S, X3 is G or Y, and / or X4 is E or Q; and iii) HC-CDR3 comprising the amino acid sequence RX1DGNPYX2MDY (SEQ ID NO: 243), wherein X1 is T or A, and / or X2 is T or A; and the VL comprises i) LC-CDR1 comprising the amino acid sequence of KASQDVSTAVX1 (SEQ ID NO: 244), wherein X1 is A or V; ii) comprising the amino acid of SEQ ID NO: 117 LC-CDR2 of the sequence; and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 118. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 115 or 221.

[0315] In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) an HC- CDR1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, or comprising at most 5, 4 in the HC-CDR , 3, 2 or 1 amino acid substitution variant; and wherein VL comprises i) LC-CDR1 comprising the amino acid sequence of KASQX1VX2TX3VX4 (SEQ ID NO: 245), wherein X1 is N or D, and X2 is G or S, X3 is N or A, and / or X4 is A or V; ii) LC-CDR2 comprising the amino acid sequence of SASYRX1X2 (SEQ ID NO: 246), wherein a) X1 is F or Y, and X2 is I or T, or b) X1X2 is FI or YT; and iii) LC-CDR3 comprising the amino acid sequence QQX1X2X3X4PX5T (SEQ ID NO: 247), wherein X1X2X3X4 is YNRN or HYST, and / or X5 and I or F. In some embodiments, the LC-CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 6, 118 or 214. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO:6. In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) amino acids comprising HC-CDR1 of SEQ ID NO: 113 sequence, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 114 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 115, or comprising at most 5, 4 , 3, 2 or 1 amino acid substitution variant; and wherein VL comprises i) LC-CDR1 comprising the amino acid sequence of KASQX1VX2TX3VX4 (SEQ ID NO: 245), wherein X1 is N or D, and X2 is G or S, X3 is N or A, and / or X4 is A or V; ii) LC-CDR2 comprising the amino acid sequence of SASYRX1X2 (SEQ ID NO: 246), wherein a) X1 is F or Y, and X2 is I or T, or b) X1X2 is FI or YT; and iii) LC-CDR3 comprising the amino acid sequence QQX1X2X3X4PX5T (SEQ ID NO: 247), wherein X1X2X3X4 is YNRN or HYST, and / or X5 and I or F. In some embodiments, the LC-CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 6, 118 or 214. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 118.

[0317] In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) amino acids comprising HC-CDR1 of SEQ ID NO: 209 sequence, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 210 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 211, or comprising at most 5, 4 , 3, 2 or 1 amino acid substitution variant; and wherein VL comprises i) LC-CDR1 comprising the amino acid sequence of KASQX1VX2TX3VX4 (SEQ ID NO: 245), wherein X1 is N or D, and X2 is G or S, X3 is N or A, and / or X4 is A or V; ii) LC-CDR2 comprising the amino acid sequence of SASYRX1X2 (SEQ ID NO: 246), wherein a) X1 is F or Y, and X2 is I or T, or b) X1X2 is FI or YT; and iii) LC-CDR3 comprising the amino acid sequence QQX1X2X3X4PX5T (SEQ ID NO: 247), wherein X1X2X3X4 is YNRN or HYST, and / or X5 and I or F. In some embodiments, the LC-CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 6, 118 or 214. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 214. In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) amino acids comprising HC-CDR1 of SEQ ID NO: 17 sequence, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, or comprising at most 5, 4 , 3, 2 or 1 amino acid substitution variant; and wherein VL comprises i) LC-CDR1 comprising the amino acid sequence of X1ASQSVX2X3X4X5X6SYMX7 (SEQ ID NO: 248), wherein X1 is K or R, and X2X3X4X5X6 is DYAGD or STSSY, and / or X7 is N or H; ii) LC-CDR2 comprising the amino acid sequence of X1ASNLES (SEQ ID NO: 249), wherein X1 is A or Y; and iii) comprising the amino acid sequence QX1X2X3X4X5PX6T ( LC-CDR3 of SEQ ID NO: 250), wherein X1X2X3X4X5 is QTNED or HSWEI, and / or X6 is R or F. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 22 or 54. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 22. In some embodiments, the antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) amino acids comprising HC-CDR1 of SEQ ID NO: 49 sequence, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 50 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 51, or comprising at most 5, 4 , 3, 2 or 1 amino acid substitution variant; and wherein VL comprises i) LC-CDR1 comprising the amino acid sequence of X1ASQSVX2X3X4X5X6SYMX7 (SEQ ID NO: 248), wherein X1 is K or R, and X2X3X4X5X6 is DYAGD or STSSY, and / or X7 is N or H; ii) LC-CDR2 comprising the amino acid sequence of X1ASNLES (SEQ ID NO: 249), wherein X1 is A or Y; and iii) comprising QX1X2X3X4X5PX6T (SEQ ID NO: 250) of the LC-CDR3 amino acid sequence, wherein X1X2X3X4X5 is QTNED or HSWEI, and / or X6 is R or F. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO: 22 or 54. In some embodiments, LC-CDR3 comprises the amino acid sequence shown in SEQ ID NO:54.

[0320] In some embodiments, the construct comprises or is an antibody or antigen-binding fragment thereof selected from the group consisting of full-length antibodies, bispecific antibodies, single-chain Fv (scFv) fragments, Fab fragments, Fab' Fragment, F(ab')2, Fv fragment, disulfide bond stabilized Fv fragment (dsFv), (dsFv)2, VHH, Fv-Fc fusion, scFv-Fc fusion, scFv-Fv fusion, bifunctional Antibodies, trifunctional antibodies and tetrafunctional antibodies.

[0321] In some embodiments, the anti-CD93 antibody portion is a full length antibody.

[0322] In some embodiments, the anti-CD93 antibody portion is a scFv.

[0323] In some embodiments, the aforementioned anti-CD93 antibody portion comprises an Fc fragment of an immunoglobulin selected from the group consisting of IgG, IgA, IgD, IgE, IgM, and combinations and hybrids thereof. In some embodiments, the aforementioned partial or full-length anti-CD93 antibody comprises an Fc fragment of an immunoglobulin selected from the group consisting of IgG1, IgG2, IgG3, IgG4, and combinations and hybrids thereof. In some embodiments, the Fc fragment has reduced effector functions compared to a corresponding wild-type Fc fragment. In some embodiments, the Fc fragment has enhanced effector functions compared to a corresponding wild-type Fc fragment. In some embodiments, the Fc fragment is altered to increase serum half-life compared to a corresponding wild-type Fc fragment. In some embodiments, the Fc fragment is altered to reduce serum half-life compared to a corresponding wild-type Fc fragment.

[0324] In some embodiments, the antibody portion comprises a humanized antibody of any of the antibody portions described herein.

[0325] In some embodiments, the anti-CD93 construct comprises or is an anti-CD93 fusion protein.

[0326] In some embodiments, an anti-CD93 construct comprises or is a multispecific anti-CD93 construct (such as a bispecific antibody).

[0327] In some embodiments, an anti-CD93 construct comprises or is an anti-CD93 immunoconjugate.

[0328] In some embodiments, the anti-CD93 construct blocks the binding of CD93 to IGFBP7. In some embodiments, the IGFBP7 is human IGFBP7. In some embodiments, the binding of CD93 to IGFBP7 is blocked by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, following preincubation of the anti-CD93 antibody with CD93 or CD93-expressing cells. 80%, 90% or more. In some embodiments, the doses of anti-CD93 antibody and CD93 are in a ratio of about 1:10, 1:6, 1:3, 1:1.5, 1:1, 4:3, 2:1, or 5:1. In some embodiments, at a concentration of about 50 µg / ml, 25 µg / ml, 10 µg / ml, 5 µg / ml, 2 µg / ml, 1 µg / ml, 0.8 µg / ml, 0.6 µg / ml or After pre-incubation with 0.4 μg / ml anti-CD93 antibody, the binding of CD93 and IGFBP7 was blocked by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more.

[0329] In some embodiments, the anti-CD93 construct blocks the binding of CD93 to MMRN2. In some embodiments, MMRN2 is human MMRN2. In some embodiments, MMRN2 is a fragment of MMRN2495-674. In some embodiments, CD93 binding to MMRN2 is at least blocked by 10%, 20%, 30%, 40%, 50%, 60%, 70%, following preincubation of the anti-CD93 antibody with CD93 or CD93-expressing cells. 80%, 90% or more. In some embodiments, the anti-CD93 construct does not block the binding of CD93 to MMRN2.

[0330] In some embodiments, the anti-CD93 construct blocks the binding of CD93 to both IGFBP7 and MMRN2.

[0331] In some embodiments, the anti-CD93 construct does not block the interaction between CD93 and IGFBP7. In some embodiments, the anti-CD93 construct does not block the interaction between CD93 and MMRN2. In some embodiments, the anti-CD93 constructs do not block the interaction between IGFBP7 or MMRN2.

[0332] In some embodiments, the CD93 is human CD93. a) Antibody affinity

[0333] The binding specificity of an antibody portion can be determined experimentally by methods known in the art. Such methods include, but are not limited to, Western blotting, ELISA, RIA-, ECL-, IRMA-, EIA-, BLI-, BIACORE™ tests, flow cytometry, and peptide scanning.

[0334] In some embodiments, the KD for binding between the antibody portion and CD93 is about 10-7M to about 10-12M, about 10-7M to about 10-8M, about 10-8M to about 10-9M, About 10-9M to about 10-10M, about 10-10M to about 10-11M, about 10-11M to about 10-12M, about 10-7M to about 10-12M, about 10-8M to about 10-12M, About 10-9M to about 10-12M, about 10-10M to about 10-12M, about 10-7M to about 10-11M, about 10-8M to about 10-11M, about 10-9M to about 10-11M, About 10-7M to about 10-10M, about 10-8M to about 10-10M, or about 10-7M to about 10-9M. In some embodiments, the KD for binding between the antibody portion and CD93 is greater than about any of 10-7M, 10-8M, 10-9M, 10-10M, 10-11M, or 10-12M. In some embodiments, CD93 is human CD93.

[0335] In some embodiments, the K association of binding between the antibody portion and CD93 is from about 103 M-1s-1 to about 108 M-1s-1, from about 103 M-1s-1 to about 104 M-1s-1, About 104M-1s-1 to about 105M-1s-1, about 105M-1s-1 to about 106M-1s-1, about 106M-1s-1 to about 107M-1s-1 or about 107M-1s-1 to about 108M-1s-1. In some embodiments, the K association of binding between the antibody portion and CD93 is about 103 M-1s-1 to about 105 M-1s-1, about 104 M-1s-1 to about 106 M-1s-1, about 105 M-1s-1 1s-1 to about 107M-1s-1, about 106M-1s-1 to about 108M-1s-1, about 104M-1s-1 to about 107M-1s-1, or about 105M-1s-1 to about 108M-1s -1. In some embodiments, the K association of binding between the antibody portion and CD93 is no more than about 103M-1s-1, 104M-1s-1, 105M-1s-1, 106M-1s-1, 107M-1s-1 or any of 108M-1s-1. In some embodiments, CD93 is human CD93.

[0336] In some embodiments, the K dissociation of the binding between the antibody portion and CD93 is about 1 s to about 10 s, about 1 to about 10 s, about 10 - 2s-1 to about 10-3s-1, about 10-3s-1 to about 10-4s-1, about 10-4s-1 to about 10-5s-1, about 10-5s-1 to about 10- 6s-1, about 1 s-1 to about 10-5s-1, about 10-2s-1 to about 10-6s-1, about 10-3s-1 to about 10-6s-1, about 10-4s- 1 to about 10-6s-1, about 10-2s-1 to about 10-5s-1, or about 10-3s-1 to about 10-5s-1. In some embodiments, the K dissociation of the binding between the antibody portion and CD93 is at least about 1 s-1, 10-2 s-1, 10-3 s-1, 10-4 s-1, 10-5 s-1 or Any of 10 -6s-1. In some embodiments, CD93 is human CD93.

[0337] In some embodiments, the anti-CD93 antibody portion or anti-CD93 construct has a higher binding affinity than (eg, has a smaller KD value) an existing anti-CD93 antibody (eg, an anti-human CD93 antibody, eg, MM01). b) chimeric or humanized antibodies

[0338] In some embodiments, the anti-CD93 antibody portion is a chimeric antibody. Certain chimeric antibodies are described, for example, in US Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In some embodiments, chimeric antibodies comprise non-human variable regions (eg, variable regions derived from mouse) and human constant regions. In some embodiments, chimeric antibodies are "class-switched" antibodies, in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0339] In some embodiments, the anti-CD93 antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce immunogenicity to humans while retaining the specificity and affinity of the parental non-human antibody. In general, humanized antibodies comprise one or more variable domains in which HVRs (eg, CDRs) (or portions thereof) are derived from non-human antibodies and FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody optionally will also comprise at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (eg, the antibody from which the HVR residues are derived), eg, to restore or improve antibody specificity or affinity.

[0340] Humanized antibodies and methods of making them are reviewed, e.g., in Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and further described, e.g., in Riechmann et al., Nature 332:323-329 (1988); Queen et al. People, Proc. Nat'l Acad. Sci. USA86:10029-10033 (1989); US Patent No. 5,821,337, No. 7,527,791, No. 6,982,321 and No. 7,087,409; 2005) (describing SDR (a-CDR) transplantation); Padlan, Mol. Immunol.28:489-498 (1991) (describing "resurfacing"); Dall'Acqua et al., Methods 36:43-60 (2005) (describing "FR shuffling"); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the "guided selection" method for FR shuffling ).

[0341] Human framework regions that can be used for humanization include, but are not limited to, framework regions selected using "best fit" methods (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); derived from Framework regions of the consensus sequence of human antibodies with a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatic mutation) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and derived from FR libraries are screened for framework regions (see eg Baca et al., J. Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem. 271:22611-22618 (1996)).

[0342] It will be appreciated that humanization of mouse-derived antibodies is a common and commonly used technique. It is thus understood that any and all humanized formats of anti-CD93 antibodies disclosed in the Sequence Listing may be used in a preclinical or clinical setting. Where any reference to a humanized format of an anti-CD93 antibody or antigen binding region thereof is used in such a preclinical or clinical setting, the humanized format is expected to have the same or similar biological activity as the original non-humanized format and profile. c) Human Antibodies

[0343] In some embodiments, the anti-CD93 antibody portion is a human antibody (referred to as a human domain antibody or a human DAb). Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr. Opin. Pharmacol.5: 368-74 (2001), Lonberg, Curr. Opin. Immunol.20:450-459 (2008), and Chen, Mol. Immunol .47(4):912-21 (2010). Transgenic mice or rats capable of producing fully human single domain antibodies (or DAbs) are known in the art. See eg US20090307787A1, US Patent No. 8,754,287, US20150289489A1, US20100122358A1 and WO2004049794.

[0344] Human antibodies (human DAbs) can be prepared by administering an immunogen to transgenic animals modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic challenge. Such animals typically contain all or a portion of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, either present extrachromosomally or integrated randomly into the animal's chromosomes. In such transgenic mice, the endogenous immunoglobulin loci are generally inactivated. For a review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). See also, for example, U.S. Patent Nos. 6,075,181 and 6,150,584, which describe XENOMOUSE™ technology; U.S. Patent No. 5,770,429, which describes HUMAB® technology; U.S. Patent No. 7,041,870, which describes K-M MOUSE® technology; and U.S. Patent Application Publication No. No. US 2007 / 0061900, which describes the VELOCIMOUSE® technology). The human variable regions of intact antibodies produced by such animals can be further modified, for example, by combining with different human constant regions.

[0345] Human antibodies (eg, human DAbs) can also be made by fusionoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines have been described for the production of human monoclonal antibodies (see, e.g., Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. pp. 51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991)). Human antibodies produced via human B cell fusion tumor technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing production of monoclonal human IgM antibodies from fusionoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human fusionomas) describe their methods. Human fusion tumor technology (three-source fusion tumor technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3 ): 185-91 (2005).

[0346] Human antibodies (eg, human DAbs) can also be produced by isolating Fv clonal variable domain sequences selected from phage display libraries of human origin. Such variable domain sequences can then be combined with the desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below. d) Library-derived antibodies

[0347] Anti-CD93 antibody portions described herein can be isolated by screening combinatorial libraries for antibodies having the desired activity. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with desired binding characteristics. Such methods are reviewed, e.g., in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (eds. O'Brien et al., Human Press, Totowa, NJ, 2001) and further described, e.g., in McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Marks and Bradbury, Methods in Molecular Biology 248:161-175 (Lo eds, Human Press , Totowa, NJ, 2003); Sidhu et al., J. Mol. Biol.338(2): 299-310 (2004); Lee et al., J. Mol. Biol.340(5): 1073-1093 (2004 ); Fellouse, Proc. Natl. Acad. Sci. USA101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004). Methods for constructing single domain antibody libraries have been described, eg see US Patent No. 7371849.

[0348] In certain phage display methods, repertoires of VH and VL genes are cloned separately by polymerase chain reaction (PCR) and randomly recombined in a phage library, which can then be analyzed as described in Winter et al., Ann. Rev. Screening was performed against antigen-binding phage as described in Immunol. 12:433-455 (1994). Phage typically present antibody fragments as scFv fragments or Fab fragments. Libraries from immunized sources provide high affinity antibodies against the immunogen without the need for construction of fusionomas. Alternatively, the original lineage (e.g. from humans) can be bred to provide a single source of antibodies against various non-self as well as self-antigens without any immunization as in Griffiths et al., EMBO J, 12: 725-734 (1993 ) mentioned. Finally, the primary library can also be prepared synthetically by cloning unrearranged V gene segments from stem cells and using PCR primers containing random sequences encoding the highly variable CDR3 region and rearranging in vitro, as in Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: U.S. Patent No. 5,750,373 and U.S. Patent Publication Nos. 2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598 No. 2007 / 0237764, 2007 / 0292936 and 2009 / 0002360.

[0349] Antibodies or antibody fragments isolated from human antibody libraries are considered human antibodies or human antibody fragments herein. e) Substitutions, insertions, deletions and variants

[0350] In some embodiments, antibody variants having one or more amino acid substitutions are provided. Relevant substitution-type mutagenic sites include HVR (CDR) and FR. Conservative substitutions are shown in Table 2 under the heading "Preferred Substitutions". More substantial variations are provided in Table 2 under the heading "Exemplary Substitutions" and are further described below when referring to amino acid side chain classes. Amino acid substitutions can be introduced into relevant antibodies and the products screened for desired activities such as retained / improved antigen binding, reduced immunogenicity, or improved ADCC or CDC. Table 2. Amino Acid Substitutions initial residue exemplary substitution better replacement Ala (A) Val; Leu; Ile Val Arg (R) Lys; Gln; Asn Lys Asn (N) Gln; His; Asp, Lys; Arg Gln Asp (D) Glu;Asn Glu Cys (C) Ser; Ala Ser Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Gly (G) Ala Ala His (H) Asn; Gln; Lys; Arg Arg Ile (I) Leu; Val; Met; Ala; Phe; Norleucine Leu Leu (L) Norleucine; Ile; Val; Met; Ala; Phe Ile Lys (K) Arg; Gln; Asn Arg Met (M) Leu; Phe; Ile Leu Phe (F) Trp; Leu; Val; Ile; Ala; Tyr Tyr Pro (P) Ala Ala Ser (S) Thr Thr Thr (T) Val; Ser Trp (W) Tyr; Phe Tyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Leu; Met; Phe; Ala; Norleucine Leu

[0351] Amino acids can be grouped according to common side chain characteristics: (1) hydrophobicity: norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilicity: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues affecting chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe.

[0352] Non-conservative substitutions would require exchanging a member of one of these classes for another class.

[0353] One type of substitutional variant involves substituting one or more hypervariable region residues of a parental antibody (eg, a humanized or human antibody). In general, the resulting variants selected for further study will have modifications (e.g., improvements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parental antibody and / or will substantially retain certain aspects of the parental antibody. some biological properties. An exemplary substitutional variant is an affinity matured antibody, which can be conveniently generated, for example, using phage display-based affinity maturation techniques such as those described herein. Briefly, one or more HVR residues are mutated and variant antibodies are displayed on phage and screened for specific biological activity (eg, binding affinity).

[0354] Alterations (eg, substitutions) can be made in the HVR, eg, to improve antibody affinity. Can be located in HVR "hot spots" (i.e. residues encoded by codons that undergo high frequency mutations during the somatic maturation process) (see e.g. Chowdhury, Methods Mol. Biol. 207:179-196 (2008)) and / or or SDR (a-CDR), wherein the resulting variant VH or VL is tested for binding affinity. Affinity maturation by construction of secondary libraries and reselection from secondary libraries has been described in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (eds. O'Brien et al., Human Press, Totowa, NJ, (2001 )). In some embodiments of affinity maturation, diversity is introduced into the variable genes selected for maturation by any of a variety of methods, such as error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis . A secondary library is then generated. This library is then screened to identify any antibody variants with the desired affinity or molecular behavior. Another method of introducing diversity involves the HVR-guided approach, in which several HVR residues (eg, 4-6 residues at a time) are randomly grouped. HVR residues involved in antigen binding can be specifically identified, for example, using alanine or histidine scanning mutagenesis or modeling. HC-CDR3 and LC-CDR3 in particular are often targeted.

[0355] In some embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, so long as such alterations do not substantially reduce the ability of the antibody to bind antigen. For example, conservative changes that do not substantially reduce binding affinity (eg, conservative substitutions as provided herein) can be made in the HVR. Such changes may be outside the HVR "hot spots" or CDRs.

[0356] A suitable method for identifying residues or regions of an antibody that can be targeted for mutagenesis is called "alanine scanning mutagenesis," as described by Cunningham and Wells (1989) Science, 244:1081-1085. describe. In this method, a residue or a group of target residues (e.g. charged residues such as Arg, Asp, His, Lys, and Glu) are identified and neutralized or negatively charged amino acids (e.g. alanine or polyalanine) are identified. ) to determine whether the interaction between the antibody and the antigen is affected. Further substitutions can be introduced at amino acid positions exhibiting functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the antigen-antibody complex is used to identify contact points between the antibody and antigen. Such contact residues and neighboring residues can be targeted or excluded as candidates for substitution. Variants can be screened to determine whether they contain the desired properties of the antibody.

[0357] Amino acid sequence insertions include amino and / or carboxyl terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as single or multiple amino acid residues inserted in the sequence. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertional variants of the antibody molecule include fusions of the N- or C-terminus of the antibody to an enzyme (eg, for ADEPT) or a polypeptide that extends the serum half-life of the antibody. f) Glycosylation variants

[0358] In some embodiments, portions of the anti-CD93 antibody are altered to increase or decrease the degree of glycosylation of the construct. Addition of glycosylation sites to or deletion of glycosylation sites from an antibody is preferably accomplished by altering the amino acid sequence to create or remove one or more glycosylation sites.

[0359] Where the antibody portion comprises an Fc region, the carbohydrate attached thereto may be altered. Native antibodies produced by mammalian cells typically comprise branched biantennary oligosaccharides, usually N-bonded to Asn297 of the CH2 domain of the Fc region. See, eg, Wright et al. TIBTECH 15:26-32 (1997). Oligosaccharides can include various carbohydrates such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as trehalose attached to GlcNAc in the "backbone" of the biantennary oligosaccharide structure . In some embodiments, the oligosaccharides in the antibody portion can be modified in order to create antibody variants with certain improved properties.

[0360] In some embodiments, the anti-CD93 antibody portion has a carbohydrate structure that lacks fucose attached (directly or indirectly) to the Fc region. For example, the amount of fucose in such antibodies may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose was calculated at Asn297 within the sugar chain relative to the sum of all sugar structures attached to Asn 297 (e.g. complex, promiscuous and high mannose structures) as measured by MALDI-TOF mass spectrometry Determine the average amount of fucose in , as described, for example, in WO 2008 / 077546. Asn297 refers to the asparagine residue located at approximately position 297 in the Fc region (EU numbering for Fc region residues); however, due to minor sequence variations in antibodies, Asn297 can also be located approximately ± ± 3 amino acids, ie between positions 294 and 300. Such fucosylation variants may have improved ADCC function. See, eg, US Patent Publication Nos. US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications on "defucosylated" or "fucose-deficient" antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US ​​2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; 586; WO 2005 / 035778; WO 2005 / 053742 ; WO2002 / 031140; Okazaki et al. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004). Examples of cell lines capable of producing afucosylated antibodies include Lec13 CHO cells lacking protein fucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); U.S. Patent Application No. US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially in Example 11) and knockout cell lines such as the alpha-1,6-fucosyltransferase gene (FUT8) Deletion of CHO cells (see e.g. Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107 ).

[0361] In some embodiments, the anti-CD93 antibody portion has bisected oligosaccharides, eg, wherein the biantennary oligosaccharides attached to the Fc region of the antibody are bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function. Examples of such antibody variants are described, eg, in WO 2003 / 011878 (Jean-Mairet et al); US Patent No. 6,602,684 (Umana et al); and US 2005 / 0123546 (Umana et al). Also provided are antibody variants in which at least one galactose residue in the oligosaccharide is attached to the Fc region. Such antibody variants may have improved CDC function. Such antibody variants are described, eg, in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.). g) Fc region variants

[0362] In some embodiments, the anti-CD93 antibody portion comprises an Fc fragment.

[0363] The term "Fc region", "Fc domain", "Fc fragment" or "Fc" refers to the C-terminal non-antigen binding region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native Fc regions as well as variant Fc regions. In some embodiments, the human IgG heavy chain Fc region extends from Cys226 to the carboxy-terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not exist without affecting the structure or stability of the Fc region. Unless otherwise stated herein, amino acid residues in an IgG or Fc region are numbered according to the EU numbering system for antibodies (also known as the EU Index), as described in: Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0364] In some embodiments, the Fc fragment is from an immunoglobulin selected from the group consisting of IgG, IgA, IgD, IgE, IgM, and combinations and hybrids thereof. In some embodiments, the Fc fragment is from an immunoglobulin selected from the group consisting of IgGl, IgG2, IgG3, IgG4, and combinations and hybrids thereof.

[0365] In some embodiments, the Fc fragment has reduced effector function compared to a corresponding wild-type Fc fragment (such as at least about 30%, 40% reduced effector function as measured by the level of antibody-dependent cellular cytotoxicity (ADCC). %, 50%, 60%, 70%, 80%, 85%, 90% or 95%).

[0366] In some embodiments, the Fc fragment is an IgG1 Fc fragment. In some embodiments, the IgG1 Fc fragment comprises the L234A mutation and / or the L235A mutation. In some embodiments, the Fc fragment is an IgG2 or IgG4 Fc fragment. In some embodiments, the Fc fragment is an IgG4 Fc fragment comprising S228P, F234A and / or L235A mutations. In some embodiments, the Fc fragment comprises the N297A mutation. In some embodiments, the Fc fragment comprises the N297G mutation.

[0367] In some embodiments, one or more amino acid modifications can be introduced into the Fc region of an antibody portion, thereby generating Fc region variants. Fc region variants may comprise human Fc region sequences (eg, human IgGl, IgG2, IgG3 or IgG4 Fc regions) comprising amino acid modifications (eg, substitutions) at one or more amino acid positions.

[0368] In some embodiments, the Fc fragment possesses some, but not all, effector functions, making it critical for partial half-life of the antibody in vivo, while certain effector functions (such as complement and ADCC) are unnecessary or detrimental Ideal candidates for applications. In vitro and / or in vivo cytotoxicity assays can be performed to confirm reduction / depletion of CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks FcγR binding (and thus likely lacks ADCC activity), but retains FcRn binding ability. Primary cells for mediating ADCC (ie, NK cells) express FcyRIII only, whereas monocytes express FcyRI, FcyRII and FcyRIII. FcR expression on hematopoietic cells is summarized in Table 2 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of related molecules are described in U.S. Patent No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986 )) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA82:1499-1502 (1985); No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med.166:1351-1361 (1987)). Alternatively, non-radioactive assays can be used (see, e.g., the ACTI™ Non-Radioactive Cytotoxicity Assay for Flow Cytometry (Cell Technology, Inc. Mountain View, CA); and the CytoTox 96® Non-Radioactive Cytotoxicity Assay (Promega, Madison, WI)). Suitable effector cells for such assays include peripheral blood mononuclear cells (PBMC) and natural killer (NK) cells. Alternatively or additionally, ADCC activity of the molecule of interest may be assessed in vivo, eg, in an animal model such as that disclosed in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). Clq binding assays can also be performed to confirm that the antibody is unable to bind Clq and thus lacks CDC activity. See eg C1q and C3c binding ELISAs in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, CDC assays can be performed (see, e.g., Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, M.S. et al., Blood 101:1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half-life can also be determined using methods known in the art (see, eg, Petkova, S.B. et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0369] Antibodies with reduced effector function include antibodies having substitutions of one or more of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (US Patent No. 6,737,056). Such Fc mutants include Fc mutants having substitutions of two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called "DANA" in which residues 265 and 297 are substituted with alanine. Fc mutants (US Patent No. 7,332,581). In some embodiments, the Fc fragment comprises the N297A mutation. In some embodiments, the Fc fragment comprises the N297G mutation.

[0370] Certain antibody variants with increased or decreased binding to FcRs are described. (See eg, US Patent No. 6,737,056; WO 2004 / 056312; and Shields et al., J. Biol. Chem. 9(2): 6591-6604 (2001).)

[0371] In some embodiments, the Fc fragment is an IgG1 Fc fragment. In some embodiments, the IgG1 Fc fragment comprises the L234A mutation and / or the L235A mutation. In some embodiments, the IgG1 Fc fragment comprises a L235A mutation and / or a G237A mutation. In some embodiments, the Fc fragment is an IgG2 or IgG4 Fc fragment. In some embodiments, the Fc fragment is an IgG4 Fc fragment comprising S228P, F234A and / or L235A mutations.

[0372] In some embodiments, the antibody portion comprises an Fc region with one or more amino acid substitutions that improve ADCC, such as substitutions at positions 298, 333, and / or 334 (EU numbering of residues) of the Fc region .

[0373] In some embodiments, changes are made in the Fc region that result in altered (i.e., improved or reduced) Clq binding and / or complement-dependent cytotoxicity (CDC), e.g., as in U.S. Pat. No. 6,194,551, WO 99 / 51642 and described in Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0374] In some embodiments, antibody partial variants comprise a variant Fc region comprising one or more amino acid substitutions that alter half-life and / or alter binding to the neonatal Fc receptor (FcRn) . Antibodies with increased half-life and improved binding to the neonatal Fc receptor (FcRn) responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24 :249 (1994)) is described in US2005 / 0014934A1 (Hinton et al.). These antibodies comprise an Fc region with one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include Fc variants having substitutions at one or more of the Fc region residues, for example a substitution of Fc region residue 434 (US Patent No. 7,371,826).

[0375] For additional examples of Fc region variants, see also Duncan and Winter, Nature 322:738-40 (1988); US Patent No. 5,648,260; US Patent No. 5,624,821; and WO 94 / 29351. h) Antibody variants engineered with cysteine

[0376] In some embodiments, it may be desirable to generate cysteine-engineered antibody portions, such as "thioMAbs," in which one or more residues of the antibody are substituted with cysteine ​​residues. In particular embodiments, the substituted residue is present at an accessible site of the antibody. By substituting these residues with cysteines, reactive thiol groups are then positioned at accessible sites of the antibody and can be used to conjugate the antibody to other moieties such as drug moieties or linker-drug moieties to produce e.g. Immunoconjugates as further described herein. In some embodiments, any one or more of the following residues may be substituted with cysteine: A118 (EU numbering) of the heavy chain; and S400 (EU numbering) of the Fc region of the heavy chain. Cysteine ​​engineered antibody portions can be produced as described, eg, in US Patent No. 7,521,541. i) Antibody Derivatives

[0377] In some embodiments, the antibody moieties described herein can be further modified to contain additional non-proteinaceous moieties known in the art and readily available. Moieties suitable for derivatization of antibodies include, but are not limited to, water soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol / propylene glycol copolymers, carboxymethylcellulose, polydextrose, polyvinyl alcohol, polyvinylpyrrolidone, Poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer) and Polydextrose or poly(n-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, polypropylene oxide / ethylene oxide copolymer, polyoxyethylated polyols (eg glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde can be advantageous in manufacturing because of its stability in water. The polymers can be of any molecular weight and can be branched or unbranched. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the number and / or type of polymers used for derivatization can be based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used for diagnosis in a given disorder, etc. to make sure.

[0378] In some embodiments, the antibody portion can be further modified to comprise one or more biologically active proteins, polypeptides or fragments thereof. "Bioactive" or "biologically active" as used interchangeably herein means the display of biological activity in vivo that performs a specific function. For example, it can mean combining with specific biomolecules, such as proteins, DNA, etc., and subsequently promoting or inhibiting the activity of such biomolecules. In some embodiments, biologically active proteins or fragments thereof include proteins and polypeptides administered to patients as active drug substances for the prevention or treatment of a disease or condition, as well as proteins and polypeptides used for diagnostic purposes, such as in diagnostic tests or enzymes analyzed in vitro, and proteins and polypeptides administered to patients to prevent disease, such as vaccines. Multispecific anti-CD93 constructs

[0379] In some embodiments, the anti-CD93 construct comprises a multispecific (e.g., bispecific) anti-CD93 construct comprising an anti-CD93 antibody moiety according to any of the anti-CD93 antibody moieties described herein, and a second binding moiety (such as a second antibody moiety) that specifically recognizes a second antigen.

[0380] In some embodiments, a multispecific anti-CD93 molecule comprises an anti-CD93 antibody portion and a second portion (such as a second antibody portion) that specifically recognizes a second antigen.

[0381] In some embodiments, the second antigen is an immune checkpoint molecule. In some embodiments, the second antigen is PD-1 or PD-L1.

[0382] In some embodiments, the second moiety is the extracellular domain (ECD) of PD-1 or PD-L1. In some embodiments, the second moiety is a PD-L1 trap or a PD-1 trap. See, eg, Nat Commun. 2018 Jun 8;9(1):2237.

[0383] In some embodiments, the second antigen is a tumor antigen.

[0384] In some embodiments, the second antigen is an angiogenic agent. In some embodiments, the angiogenic agent is an antibody to VEGF (eg, human VEGF). In some embodiments, the angiogenic agent is a VEGF receptor. In some embodiments, the angiogenic agent is VEGFR1 (eg, human VEGFR1). In some embodiments, the angiogenic agent is VEGFR2 (eg, human VEGFR2).

[0385] In some embodiments, the second portion comprises the extracellular domain (ECD) of a VEGF receptor. In some embodiments, the second portion comprises the ECD of VEGFR1 and / or VEGFR2. In some embodiments, the second part comprises a VEGF-Trap. See, eg, Proc Natl Acad Sci USA. 2002 Aug 20;99(17):11393-8.

[0386] In some embodiments, the second antibody portion and the anti-CD93 antibody portion are fused to each other via a linker, such as any linker described herein having any operable format that allows the appropriate function of the binding portion. In some embodiments, the linker is a GS linker. In some embodiments, the linker is selected from the group consisting of SEQ ID NO: 225-232 and 338.

[0387] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 antibody moiety according to any one of the anti-CD93 antibody moieties described herein Antibody portion; b) a second antibody portion that specifically recognizes PD-L1 (anti-PD-L1 antibody portion).

[0388] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL); b) with at least one or both of the heavy chains of the anti-CD93 full-length antibody A fused anti-PD-L1 antibody portion (such as any of the antibody portions described herein). In some embodiments, anti-PD-L1 antibody portions are fused to the N-terminus of two heavy chains. In some embodiments, portions of the anti-PD-L1 antibody are fused to the C-terminus of the two heavy chains.

[0389] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-PD-L1 antibody portion comprising two heavy chains and two A full-length antibody of a light chain, in which two heavy chains each comprise a heavy chain variable region (VH) and two light chains each comprise a light chain variable region (VL); b) the same weight as the anti-PD-L1 full-length antibody An anti-CD93 antibody portion (such as any of the anti-CD93 antibody portions described herein) to which at least one or both of the chains are fused. In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two heavy chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two heavy chains.

[0390] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein Two heavy chains each comprising a heavy chain variable region (VH) and two light chains each comprising a light chain variable region (VL); b) fused to at least one or both of the light chains of an anti-CD93 full-length antibody An anti-PD-L1 antibody portion (such as any of the antibody portions described herein). In some embodiments, anti-PD-L1 antibody portions are fused to the N-terminus of two light chains. In some embodiments, anti-PD-L1 antibody portions are fused to the C-terminus of two light chains.

[0391] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-PD-L1 antibody portion comprising two heavy chains and two A full-length antibody with a light chain, in which two heavy chains each comprise a heavy chain variable region (VH) and two light chains each comprise a light chain variable region (VL); b) with the light chain of an anti-PD-L1 full-length antibody An anti-CD93 antibody portion (such as any of the antibody portions described herein) to which at least one or both of the chains are fused. In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two light chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two light chains.

[0392] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 antibody according to any one of the anti-CD93 antibody moieties described herein Antibody portion; b) a second antibody portion that specifically recognizes PD-1 (anti-PD-1 antibody portion).

[0393] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL); b) with at least one or both of the heavy chains of the anti-CD93 full-length antibody A fused anti-PD-1 antibody portion (such as any of the antibody portions described herein). In some embodiments, anti-PD-1 antibody portions are fused to the N-terminus of two heavy chains. In some embodiments, anti-PD-1 antibody portions are fused to the C-terminus of two heavy chains.

[0394] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-PD-1 antibody portion comprising two heavy chains and two A full-length light chain antibody, in which two heavy chains each comprise a heavy chain variable region (VH) and two light chains each comprise a light chain variable region (VL); b) the same weight as the anti-PD-1 full-length antibody An anti-CD93 antibody portion (such as any of the anti-CD93 antibody portions described herein) to which at least one or both of the chains are fused. In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two heavy chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two heavy chains.

[0395] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL); b) at least one or both of the light chains of the anti-CD93 full-length antibody A fused anti-PD-1 antibody portion (such as any of the antibody portions described herein). In some embodiments, anti-PD-1 antibody portions are fused to the N-terminus of two light chains. In some embodiments, anti-PD-1 antibody portions are fused to the C-terminus of two light chains.

[0396] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-PD-1 antibody portion comprising two heavy chains and two A full-length antibody with a light chain, in which two heavy chains each comprise a heavy chain variable region (VH) and two light chains each comprise a light chain variable region (VL); b) with the light chain of an anti-PD-1 full-length antibody An anti-CD93 antibody portion (such as any of the antibody portions described herein) to which at least one or both of the chains are fused. In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two light chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two light chains.

[0397] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 antibody moiety according to any one of the anti-CD93 antibody moieties described herein An antibody portion; b) a second binding portion that specifically recognizes VEGF.

[0398] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL); b) with at least one or both of the heavy chains of the anti-CD93 full-length antibody The fusion specifically recognizes the second binding portion of VEGF. In some embodiments, the second binding moiety is fused to the N-terminus of the two heavy chains. In some embodiments, the second binding moiety is fused to the C-terminus of both heavy chains.

[0399] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-VEGF antibody portion comprising two heavy chains and two light chains The full-length antibody of wherein two heavy chains each comprise heavy chain variable region (VH) and two light chains each comprise light chain variable region (VL); b) with at least one of the heavy chains of anti-VEGF full-length antibody One or both fused anti-CD93 antibody portions (such as any of the anti-CD93 antibody portions described herein). In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two heavy chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two heavy chains.

[0400] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL); b) at least one or both of the light chains of the anti-CD93 full-length antibody The fusion specifically recognizes the second binding portion of VEGF. In some embodiments, the second binding moiety is fused to the N-terminus of the two light chains. In some embodiments, a second binding moiety that specifically recognizes VEGF is fused to the C-termini of the two light chains.

[0401] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-VEGF antibody portion comprising two heavy chains and two light chains The full-length antibody of wherein two heavy chains each comprise heavy chain variable region (VH) and two light chains each comprise light chain variable region (VL); b) with at least one of the light chains of anti-VEGF full-length antibody Anti-CD93 antibody portions (such as any of the antibody portions described herein) fused to one or both. In some embodiments, anti-CD93 antibody portions are fused to the N-terminus of two light chains. In some embodiments, anti-CD93 antibody portions are fused to the C-terminus of two light chains.

[0402] In some embodiments, there is provided an anti-CD93 construct comprising: a) a full-length antibody that specifically recognizes CD93 comprising two heavy chains and two light chains, wherein each of the two heavy chains comprises a heavy Chain variable region (VH), this heavy chain variable region comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and comprising SEQ ID The HC-CDR3 of the amino acid sequence of NO: 291, and wherein each of the two light chains comprises a light chain variable region (VL), and the light chain variable region comprises an LC comprising the amino acid sequence of SEQ ID NO: 292 -CDR1, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294, and b) comprising the amino acid sequence of SEQ ID NO: 325 A VEGF-binding portion, wherein the VEGF-binding portion is fused to one or both of the heavy chains of the full-length antibody. In some embodiments, the VEGF binding portion is fused to the C-terminus of the two heavy chains of the full length antibody. In some embodiments, the VEGF binding portion is fused to the full length antibody without a linker. In some embodiments, the VEGF binding portion is fused to the full length antibody via a linker. In some embodiments, the linker is a GS linker or is selected from the group consisting of SEQ ID NOs: 225-232 and 338. In some embodiments, the linker comprises the amino acid sequence of SEQ ID NO: 338. In some embodiments, the anti-CD93 VH comprises the amino acid sequence of any one of SEQ ID NO: 287 and 319-321, or comprises at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% in any one of the amino acid sequence variants of sequence identity; and the VL comprises any one of SEQ ID NO: 288 and 322-324 or comprising at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity Variants of amino acid sequences. In some embodiments, the full length antibody has an IgGl isotype (such as a human IgGl isotype). In some embodiments, the heavy chain comprises the amino acid sequence of SEQ ID NO: 342, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). % or 99%) sequence identity variants of amino acid sequences. In some embodiments, the light chain comprises the amino acid sequence of SEQ ID NO: 343, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). % or 99%) sequence identity variants of amino acid sequences. In some embodiments, there is provided an anti-CD93 construct comprising: a heavy chain fusion polypeptide comprising the amino acid sequence shown in SEQ ID NO: 366, or comprising at least about 80% ( Variants of amino acid sequences, such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity; and light chains, the light chains The chain comprises the amino acid sequence shown in SEQ ID NO: 367, or comprises at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) A variant of the amino acid sequence of any one of the sequence identity.

[0403] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and iii) ) comprising the HC-CDR3 of the amino acid sequence of SEQ ID NO: 3, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and the anti-CD93 VL comprising i ) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) LC comprising the amino acid sequence of SEQ ID NO: 6 - CDR3, or a variant thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0404] In some embodiments, the anti-CD93 VH comprises i) an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 or 304, ii) an HC comprising the amino acid sequence of SEQ ID NO: 18 or 305 -CDR2 and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and resistant to CD93 VL comprising i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, 301, 302, 303 or 306, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising LC-CDR3 of the amino acid sequence of SEQ ID NO: 22, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0405] In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290, and iii ) comprising the HC-CDR3 of the amino acid sequence of SEQ ID NO: 291, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and the anti-CD93 VL comprising i ) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) LC comprising the amino acid sequence of SEQ ID NO: 294 - CDR3, or a variant thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR. In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 295, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 296 and iii) comprising SEQ ID NO: 296 HC-CDR3 of the amino acid sequence of ID NO: 297, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and the anti-CD93 VL comprises i) comprising SEQ ID NO: LC-CDR1 of the amino acid sequence of 298, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 299 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 300, Or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0406] In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of this application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application. Exemplary Anti-PD-L1 Antibody Portions

[0407] Exemplary anti-PD-L1 antibody portions include, but are not limited to, those anti-PD-L1 antibody portions described in WO2019228514A1, WO2019227490A1, and WO2020019232A1.

[0408] In some embodiments, the anti-PD-L1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) Part, wherein the antibody part competes with an antibody or antibody fragment comprising a second heavy chain variable region (VH-2) and a second light chain variable region (VL-2) for binding epitopes of PD-L1, wherein the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 251, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 252, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 253 CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 254, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 255 and comprising the amino acid of SEQ ID NO: 256 Sequence of LC-CDR3.

[0409] In some embodiments, the anti-PD-L1 moiety comprises an HC comprising an amino acid sequence of CDR1, CDR2, and CDR3, respectively, within the VH chain region having the sequence set forth in SEQ ID NO: 281, 282, or 283 -CDR1, HC-CDR2, and HC-CDR3; and LC-CDR1, respectively comprising the amino acid sequences of CDR1, CDR2, and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 284, 285, or 286, LC-CDR2 and LC-CDR3.

[0410] In some embodiments, the anti-PD-L1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) VH includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 251, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 252, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 253 CDR3, or variants thereof comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and b) VL comprising an LC- comprising the amino acid sequence of SEQ ID NO: 254 CDR1, LC-CDR2 comprising any one of the amino acid sequences of SEQ ID NO: 255 and LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 256, or LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 256 Variants comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in .

[0411] In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0412] In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 281, 282 or 283, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%). %, 97%, 98% or 99% in any one of 99%) amino acid sequence variants of sequence identity; and VL comprises the amino acid sequence of SEQ ID NO: 284, 285 or 286, or comprises a A variant of an amino acid sequence having about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 281, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 284, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 282, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 285, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 283, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 286, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0413] In some embodiments, the second antibody portion and the anti-CD93 antibody portion are fused to each other via a linker, such as any linker described herein having any operable format that allows for the appropriate function of the binding portion. Exemplary Anti-PD-1 Antibody Portions

[0414] Exemplary anti-PD-1 antibody portions include, but are not limited to, those described in WO2018133842 and WO2018133837.

[0415] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) Part, wherein the antibody part competes for the binding epitope of PD-1 with an antibody or antibody fragment comprising a second heavy chain variable region (VH-2) and a second light chain variable region (VL-2), wherein the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 257, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 258, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 259 CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 260, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 261 and comprising the amino acid of SEQ ID NO: 262 Sequence of LC-CDR3.

[0416] In some embodiments, the anti-PD-1 moiety comprises HC-CDR1, HC comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 275, respectively. -CDR2 and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 276, respectively.

[0417] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) VH includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 257, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 258, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 259 CDR3, or variants thereof comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and b) VL comprising an LC- comprising the amino acid sequence of SEQ ID NO: 260 CDR1, LC-CDR2 comprising any one of the amino acid sequences of SEQ ID NO: 261 and LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 262, or LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 262 Variants comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in . In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0418] In some embodiments, the second antibody portion comprises a humanized antibody portion derived from a murine antibody comprising a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 275 and a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO: 276.

[0419] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) Part, wherein the antibody part competes for the binding epitope of PD-1 with an antibody or antibody fragment comprising a second heavy chain variable region (VH-2) and a second light chain variable region (VL-2), wherein the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 263, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 264, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 265 CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 266, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 267 and comprising the amino acid of SEQ ID NO: 268 Sequence of LC-CDR3.

[0420] In some embodiments, the anti-PD-1 moiety comprises HC-CDR1, HC comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 277, respectively. -CDR2 and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 respectively comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 278.

[0421] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) VH includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 263, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 264, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 265 CDR3, or variants thereof comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and b) VL comprising an LC- comprising the amino acid sequence of SEQ ID NO: 266 CDR1, LC-CDR2 comprising any one of the amino acid sequences of SEQ ID NO: 267 and LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 268, or LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 268 Variants comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in . In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application. In some embodiments, the anti-PD-1 antibody portion comprises a humanized antibody portion derived from a murine antibody comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 277 ( VH) and a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO: 278.

[0423] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises an antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL) Part, wherein the antibody part competes for the binding epitope of PD-1 with an antibody or antibody fragment comprising a second heavy chain variable region (VH-2) and a second light chain variable region (VL-2), wherein the VH-2 includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 269, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 270, and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 271 CDR3, and the VL-2 includes LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 272, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 273 and comprising the amino acid of SEQ ID NO: 274 Sequence of LC-CDR3.

[0424] In some embodiments, the anti-PD-1 moiety comprises HC-CDR1, HC comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VH chain region having the sequence shown in SEQ ID NO: 279, respectively. -CDR2 and HC-CDR3; and LC-CDR1, LC-CDR2 and LC-CDR3 comprising the amino acid sequences of CDR1, CDR2 and CDR3 in the VL chain region having the sequence shown in SEQ ID NO: 280, respectively.

[0425] In some embodiments, the anti-PD-1 antibody portion (such as scFv) used in the multispecific anti-CD93 construct comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: a) VH includes HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 269, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 270 and HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 271 CDR3, or variants thereof comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR; and b) VL comprising an LC- comprising the amino acid sequence of SEQ ID NO: 272 CDR1, LC-CDR2 comprising any one of the amino acid sequences of SEQ ID NO: 273 and LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 274, or LC-CDR3 comprising any one of the amino acid sequences of SEQ ID NO: 274 Variants comprising up to a total of about 5, 4, 3, 2 or 1 amino acid substitutions in . In some embodiments, the amino acid substitutions described above are limited to the "exemplary substitutions" shown in Table 2 of the present application. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" shown in Table 2 of this application.

[0426] In some embodiments, the second antibody portion comprises a humanized antibody portion derived from a murine antibody comprising a heavy chain variable region (VH) comprising the amino acid sequence set forth in SEQ ID NO: 279 and a light chain variable region (VL) comprising the amino acid sequence shown in SEQ ID NO: 280).

[0427] In some embodiments, the second antibody portion and the anti-CD93 antibody portion are fused to each other via a linker, such as any linker described herein having any operable format that allows for the appropriate function of the binding portion. Exemplary binding moieties that specifically recognize VEGF

[0428] Exemplary binding moieties that specifically recognize VEGF include, but are not limited to, Avastin, ramucirumab, or VEGF-Trap (aflibercept), or variants or functional portions thereof.

[0429] In some embodiments, the binding moiety that specifically recognizes VEGF used in the multispecific anti-CD93 construct is an antibody moiety (such as a scFv) comprising a heavy chain variable region (VH) and a light chain variable region. The antibody part of region (VL), wherein the VH comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 326, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 327 and comprising SEQ ID NO: 328 The HC-CDR3 of the amino acid sequence of the VL, and the VL comprises the LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 329, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 330 and the LC-CDR2 comprising the amino acid sequence of SEQ ID NO : LC-CDR3 of the amino acid sequence of 331.

[0430] In some embodiments, the binding moiety that specifically recognizes VEGF used in the multispecific anti-CD93 construct is an antibody moiety (such as a scFv) comprising a heavy chain variable region (VH) and a light chain variable domain. The antibody part of region (VL), wherein the VH comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 332, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 333 and comprising SEQ ID NO: 334 The HC-CDR3 of the amino acid sequence of the VL, and the VL comprises the LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 335, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 336 and the LC-CDR2 comprising the amino acid sequence of SEQ ID NO : LC-CDR3 of the amino acid sequence of 337.

[0431] In some embodiments, the binding moiety specifically recognizing VEGF used in the multispecific anti-CD93 construct is an antibody moiety (such as scFv) comprising a heavy chain variable region (VH) and a light chain variable region The antibody part of region (VL), wherein the VH comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and comprising SEQ ID NO: 291 The HC-CDR3 of the amino acid sequence of the VL, and the VL comprises the LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and the LC-CDR2 comprising the amino acid sequence of SEQ ID NO : LC-CDR3 of the amino acid sequence of 294.

[0432] In some embodiments, the binding moiety that specifically recognizes VEGF used in the multispecific anti-CD93 construct comprises the amino acid sequence of SEQ ID NO: 325.

[0433] In some embodiments, the anti-CD93 construct is a multispecific (e.g., bispecific) anti-CD93 construct comprising: a) an anti-CD93 full-length antibody comprising two heavy chains and two light chains, wherein the two heavy chains each comprise a heavy chain variable region (VH) and the two light chains each comprise a light chain variable region (VL), wherein the VH comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289 , HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 291, and the VL comprises LC comprising the amino acid sequence of SEQ ID NO: 292 - CDR1, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294; and b) a binding portion specifically recognizing VEGF, which binds to anti-CD93 The C-termini of the two heavy chains of the full-length antibody are fused. In some embodiments, the binding moiety that specifically recognizes VEGF used in the multispecific anti-CD93 construct comprises the amino acid sequence of SEQ ID NO: 325, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%). %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity. In one embodiment, an anti-CD93 construct is provided, comprising: a heavy chain fusion polypeptide comprising the amino acid sequence shown in SEQ ID NO: 366, or comprising at least about 80% ( Variants of amino acid sequences, such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity; and light chains, the light chains The chain comprises the amino acid sequence shown in SEQ ID NO: 367, or comprises at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) A variant of the amino acid sequence of any one of the sequence identity.

[0434] In some embodiments, the multispecific anti-CD93 construct is capable of blocking the interaction between CD93 and IGFBP7. In some embodiments, the multispecific anti-CD93 construct is capable of blocking the interaction between CD93 and IGFBP7 by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.

[0435] In some embodiments, a multispecific anti-CD93 construct is capable of blocking the interaction between CD93 and MMRN2. In some embodiments, the multispecific anti-CD93 construct is capable of blocking the interaction between CD93 and MMRN2 by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.

[0436] In some embodiments, the multispecific anti-CD93 construct is capable of measuring less than 1 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, A dissociation constant of about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, or greater than about 50 nM binds to VEGFA. In some embodiments, the multispecific anti-CD93 construct is capable of binding to VEGFA with a dissociation constant of about 2 nM as measured by biolayer interferometry. Additional anti-CD93 fusion protein

[0437] In some embodiments, an anti-CD93 construct comprises an anti-CD93 antibody portion (eg, an anti-CD93 scFv) and a second portion.

[0438] In some embodiments, the second moiety comprises a half-life extending moiety. In some embodiments, the half-life extending moiety is an albumin binding moiety (eg, an albumin binding antibody moiety). In some embodiments, the anti-CD93 antibody portion and the half-life extending portion are linked via a linker, such as any of the linkers described in the "Linkers" section.

[0439] In some embodiments, the second portion comprises the extracellular domain of the receptor. In some embodiments, the second moiety is the extracellular domain (ECD) of PD-1 or PD-L1. In some embodiments, the second moiety is a PD-L1 trap or a PD-1 trap. See, eg, Nat Commun. 2018 Jun 8;9(1):2237. In some embodiments, the second portion comprises the extracellular domain (ECD) of a VEGF receptor. In some embodiments, the second portion comprises the ECD of VEGFR1 and / or VEGFR2. In some embodiments, the second part comprises a VEGF-Trap. See, eg, Proc Natl Acad Sci USA. 2002 Aug 20;99(17):11393-8. Anti-CD93 Immunoconjugate

[0440] The application also provides anti-CD93 immunoconjugates comprising an anti-CD93 antibody moiety (such as any of the CD93 antibody moieties described herein) and a second agent. In some embodiments, the second agent is a therapeutic agent. In some embodiments, the second agent is a marker. Linker

[0441] In some embodiments, the anti-CD93 constructs described herein are separated in two parts (e.g., the anti-CD93 antibody part and the half-life extending part, the anti-CD93 antibody part and the second binding part in the multispecific constructs described above) contains one or more linkers. The length, degree of flexibility, and / or other properties of linkers used in anti-CD93 constructs may have some effect on properties including, but not limited to, affinity, specificity, or avidity for one or more particular antigens or epitopes. affect. For example, longer linkers can be chosen to ensure that two adjacent domains do not spatially interfere with each other. In some embodiments, a linker (such as a peptide linker) comprises flexible residues (such as glycine and serine) such that adjacent domains are free to move relative to each other. For example, a glycine-serine doublet may be a suitable peptide linker. In some embodiments, the linker is a non-peptide linker. In some embodiments, the linker is a peptide linker. In some embodiments, the linker is a non-cleavable linker. In some embodiments, the linker is a cleavable linker.

[0442] Other linker considerations include effects on the physical or pharmacokinetic properties of the resulting compound, such as solubility, lipophilicity, hydrophilicity, hydrophobicity, stability (more or less stable and intended to degrade), rigidity , flexibility, immunogenicity, modulation of antibody binding, ability to incorporate into micelles or liposomes, and the like. peptide linker

[0443] The peptide linker can have a naturally occurring sequence or a non-naturally occurring sequence. For example, sequences derived from the hinge region of heavy chain-only antibodies can be used as linkers. See eg WO1996 / 34103.

[0444] The peptide linker can be of any suitable length. In some embodiments, the length of the peptide linker is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, Any of 19, 20, 25, 30, 35, 40, 50, 75, 100 or more amino acids. In some embodiments, the length of the peptide linker is no more than about 100, 75, 50, 40, 35, 30, 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, Any of 9, 8, 7, 6, 5 or fewer amino acids. In some embodiments, the length of the peptide linker is any of the following: about 1 amino acid to about 10 amino acids, about 1 amino acid to about 20 amino acids, about 1 amino acid Amino acids to about 30 amino acids, about 5 amino acids to about 15 amino acids, about 10 amino acids to about 25 amino acids, about 5 amino acids to about 30 amino acids amino acids, about 10 amino acids to about 30 amino acids long, about 30 amino acids to about 50 amino acids, about 50 amino acids to about 100 amino acids, or about 1 amine amino acids to about 100 amino acids.

[0445] An essential technical feature of this type of peptide linker is that the peptide linker does not comprise any polymerization activity. The characteristics of peptide linkers (lack of facilitation including secondary structure) are known in the art and described, for example, in Dall'Acqua et al. (Biochem. (1998) 37, 9266-9273), Cheadle et al. (Mol. Immunol (1992) 29, 21-30) and Raag and Whitlow (FASEB (1995) 9(1), 73-80). In the case of "peptide linkers", a preferred amino acid is Gly. Furthermore, peptide linkers that do not promote any secondary structure are also preferred. Linkage of domains to one another can be provided by, for example, genetic engineering. Methods for making fused and operably linked bispecific single chain constructs and expressing them in mammalian cells or bacteria are well known in the art (e.g. WO 99 / 54440, Ausubel, Current Protocols in Molecular Biology, Green Publishing Associates and Wiley Interscience, N. Y. 1989 and 1994 or Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y., 2001).

[0446] The peptide linker may be a stable linker that is not cleavable by proteases, especially matrix metalloproteases (MMPs).

[0447] The linker can also be a flexible linker. Exemplary flexible linkers include glycine polymer (G)n (SEQ ID NO: 225), glycine-serine polymer (including for example (GS)n (SEQ ID NO: 226), ( GSGGS)n (SEQ ID NO: 227), (GGGGS)n (SEQ ID NO: 228) and (GGGS)n (SEQ ID NO: 229), wherein n is an integer of at least one), glycine-propylamine Acid polymers, alanine-serine polymers, and other flexible linkers known in the art. Glycine and glycine-serine polymers are relatively unstructured and are thus able to act as neutral tethers between the components. Glycine enters the φ-ψ gap even more readily than alanine, and is much less restricted than residues with longer side chains (see Scheraga, Rev. Computational Chem. 11 173-142 (1992)). Those of ordinary skill will recognize that the design of antibody fusion proteins may include a fully or partially flexible linker such that the linker may include a flexible linker portion and one or more portions that impart less flexible structures, To obtain the desired antibody fusion protein structure.

[0448] In addition, exemplary linkers also include amino acid sequences, such as (GGGGS)n (SEQ ID NO: 228), wherein n is an integer between 1 and 8, such as (GGGGS) 3 (SEQ ID NO: 230; hereinafter referred to as "(G4S)3" or "GS3") or (GGGGS)6 (SEQ ID NO: 231; hereinafter referred to as "(G4S)6" or "GS6"). In some embodiments, the peptide linker comprises the amino acid sequence (GSTSGSGKPGSGEGS)n (SEQ ID NO: 232), where n is an integer between 1 and 3. non-peptide linker

[0449] Coupling of the two moieties can be accomplished by any chemical reaction that will bind the two molecules, as long as the two components retain their respective activities, e.g. a second agent that binds to CD93 and an anti-CD93 multispecific antibody respectively. Can. This linking can involve many chemical mechanisms such as covalent binding, affinity binding, intercalation, coordinate binding and complexation. In some embodiments, the binding is covalent. Covalent attachment can be achieved by direct condensation of existing side chains or by incorporation of external bridging molecules. In this context, many divalent or multivalent linking agents are suitable for coupling protein molecules. For example, representative coupling agents can include organic compounds such as thioesters, carbodiimides, succinimidyl esters, diisocyanates, glutaraldehyde, diazobenzene, and hexamethylenediamine. This list is not intended to be an exhaustive list of the various types of couplers known in the art, but rather is an example of the more common couplers (see Killen and Lindstrom, Jour. Immun. 133:1335-2549 (1984); Jansen et al., Immunological Reviews 62:185-216 (1982); and Vitetta et al., Science 238:1098 (1987)).

[0450] Linkers that may be suitable for use in the present application are described in the literature (see, for example, Ramakrishnan, S. et al., Cancer Res. 44:201-208 (1984), which describes MBS (M-maleyl iminobenzoyl-N-hydroxybutanediimide ester)). In some embodiments, non-peptide linkers used herein include: (i) EDC (1-ethyl-3-(3-dimethylamino-propyl)carbodiimide hydrochloride; (ii) SMPT (4-butadiiminooxycarbonyl-α-methyl-α-(2-pyridyl-dithio)-toluene (Pierce Chem. Co., catalog number (21558G); (iii) SPDP ( Succimidyl-6[3-(2-pyridyldithio)propionylamino]hexanoate (Pierce Chem. Co., Cat. No. 21651G); (iv) Sulfo-LC-SPDP ( Sulfosuccimidyl 6[3-(2-pyridyldithio)-acrylamide]hexanoate (Pierce Chem. Co., Cat. No. 2165-G); and (v) conjugated to EDC Sulfo-NHS (N-Hydroxysulfo-succinimide: Pierce Chem. Co., Cat. No. 24510). In some embodiments, the linker is a PEG-containing linker.

[0451] The above-mentioned linkers contain components with different properties, so that bispecific antibodies with different physicochemical properties can be generated. For example, sulfo-NHS esters of alkyl carboxylic acids are more stable than sulfo-NHS esters of aromatic carboxylic acids. Linkers containing NHS-esters are less soluble than sulfo-NHS esters. In addition, the linker SMPT contains a sterically hindered disulfide bond and can form an antibody fusion protein with enhanced stability. Disulfide bonds are generally less stable than other bonds because disulfide bonds are cleaved in vitro, which makes antibody fusion proteins less usable. In particular, sulfo-NHS can enhance the stability of carbodiimide couplings. Carbodiimide coupling, such as EDC, when used in conjunction with sulfo-NHS, forms esters that are more resistant to hydrolysis than carbodiimide coupling alone. III. Preparation method

[0452] In some embodiments, a method of preparing an anti-CD93 construct or antibody portion that specifically binds to CD93 and compositions produced during preparation of the anti-CD93 construct or antibody portion, such as polynucleotides, Nucleic acid construct, vector, host cell or culture medium. The anti-CD93 constructs or antibody portions or compositions described herein can be prepared by a number of methods as outlined below and more specifically described in the Examples. Antibody expression and production

[0453] The antibodies described herein (including anti-CD93 monoclonal antibodies, anti-CD93 bispecific antibodies, and anti-CD93 antibody portions) can be prepared using any method known in the art, including those described below and in the Examples. monoclonal antibody From a population of substantially homogeneous antibodies (i.e., except for possible naturally occurring mutations and / or post-translational modifications (e.g., isomerization, amidation) that may be present in minor amounts, the individual individuals comprising the population Antibodies are the same) to obtain monoclonal antibodies. Thus, the modifier "monoclonal" indicates that the antibody is not characteristic of a mixture of discrete antibodies. For example, monoclonal antibodies can be produced by the fusionoma method first described by Kohler et al., Nature, 256:495 (1975), or by recombinant DNA methods (US Patent No. 4,816,567). In the fusionoma approach, a mouse or other suitable host animal (such as a hamster or llama) is immunized as described above to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization . Alternatively, lymphocytes can be immunized ex vivo. Lymphocytes are then fused with myeloma cells using a suitable fusion agent such as polyethylene glycol to form fusionoma cells (Goding, Monoclonal Antibodies: Principles and Practice, pp. 59-103 (Academic Press, 1986). On Camel See also Example 1 for immunization.

[0455] The immunizing agent will generally include the antigenic protein or a fusion variant thereof. Generally, peripheral blood lymphocytes ("PBL") are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with the immortalized cell line using a suitable fusion agent such as polyethylene glycol to form confluent tumor cells. Goding, Monoclonal Antibodies: Principles and Practice, Academic Press (1986), pp. 59-103.

[0456] Immortalized cell lines are usually transformed mammalian cells, especially myeloma cells of rodent, bovine and human origin. Typically, rat or mouse myeloma cell lines are used. The fusionoma cells thus prepared are seeded and grown in a suitable medium, which preferably contains one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells do not contain the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT), the culture medium for fusionomas will typically include hypoxanthine, aminopterin, and thymidine (HAT medium), these substances prevent the growth of cells lacking HGPRT.

[0457] Preferred immortalized myeloma cells are those that fuse efficiently, support stable high levels of antibody production by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these myeloma cell lines, preferred are murine myeloma cell lines, such as those derived from MOPC-21 and MPC-11 mouse tumors, obtained from Salk Institute Cell Distribution Center (San Diego, Calif. , USA); and SP-2 cells (and derivatives thereof, such as X63-Ag8-653), obtained from the American Type Culture Collection, Manassas, Va. USA). Human myeloma and mouse-human heteromyeloma cell lines have also been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).

[0458] The culture medium in which the fusionoma cells are growing is analyzed for the production of monoclonal antibodies to the antigen. Preferably, the binding specificity of monoclonal antibodies produced by fusion tumor cells is determined by immunoprecipitation or by in vitro binding assays such as flow cytometry, radioimmunoassay (RIA) or enzyme-linked immunosorbent assay. (ELISA)) to measure.

[0459] The medium in which the fusionoma cells are grown can be assayed for the presence of monoclonal antibodies to the antigen. Preferably, the binding affinity and specificity of monoclonal antibodies can be determined by immunoprecipitation or by in vitro binding assays such as radioimmunoassay (RIA), enzyme-linked assay (ELISA) or BLI. Such techniques and analyzes are known in the art. Binding affinity can be determined, for example, by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980).

[0460] After identification of fusionoma cells producing antibodies with the desired specificity, affinity and / or activity, clonal lines can be subcloned by limiting dilution procedures and rendered clonal by standard methods (Goding, supra). grow. Suitable media for this purpose include, for example, D-MEM or RPMI-1640 media. Additionally, fusionoma cells can be grown in vivo as tumors in mammals.

[0461] Monoclonal antibodies secreted by hypoclonal lines are suitably purified by conventional immunoglobulin procedures such as protein A-agarose, hydroxyapatite chromatography, ion exchange chromatography, gel electrophoresis, dialysis or affinity purification. And chromatographic separation from culture medium, ascitic fluid or serum.

[0462] Monoclonal antibodies can also be prepared by recombinant DNA methods, such as those described in US Patent No. 4,816,567 and those described above. mRNA encoding monoclonal antibodies can be readily isolated and sequenced using conventional procedures (eg, by using oligonucleotide probes that are capable of binding specifically to cDNA encoding the heavy and light chains of murine antibodies). Fusoma cells serve as a preferred source of such mRNA. Once isolated, the cDNA can be placed into an expression vector and subsequently transfected into host cells such as E. coli cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells (which do not otherwise produce immunoglobulins) , to produce monoclonal antibodies in such recombinant host cells. Review articles on recombinant expression in bacteria of antibody-encoding DNA include Skerra et al., Curr. Opinion in Immunol., 5:256-262 (1993) and Plückthun, Immunol. Revs. 130:151-188 (1992).

[0463] In another embodiment, antibodies can be isolated from antibody phage libraries generated using the techniques described in McCafferty et al., Nature, 348:552-554 (1990). Clackson et al., Nature, 352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991) describe the use of phage libraries to isolate murine and human antibodies, respectively. Subsequent publications describe the generation of high-affinity (nM range) human antibodies by chain shuffling (Marks et al., Bio / Technology, 10:779-783 (1992)) and combined infection and in vivo recombination as methods for constructing very large Strategies for phage libraries (Waterhouse et al., Nucl. Acids Res., 21:2265-2266 (1993)). Therefore, these techniques are viable alternatives to traditional monoclonal antibody fusion tumor techniques for isolation of monoclonal antibodies.

[0464] The DNA may also be modified, for example, by replacing the homologous murine sequences with the coding sequences of the human heavy and light chain constant domains (US Pat. No. 4,816,567; Morrison et al., Proc. Natl Acad. Sci. USA , 81:6851 (1984)) or by covalently joining all or a portion of the coding sequence of the non-immunoglobulin polypeptide to the immunoglobulin coding sequence. Typically, such non-immunoglobulin polypeptides are substituted for the constant domains of the antibody, or for the variable domains of one of the antibody's antigen-binding sites, to generate chimeric bivalent antibodies comprising a single target for the antigen An antigen-binding site with specificity and another antigen-binding site with specificity for a different antigen.

[0465] The monoclonal antibodies described herein may be monovalent, and methods for their preparation are well known in the art. For example, one method involves recombinant expression of immunoglobulin light chains and modified heavy chains. The heavy chain is typically truncated at any point in the Fc region to prevent cross-linking of the heavy chain. Alternatively, the relevant cysteine ​​residue may be substituted with another amino acid residue or deleted to prevent cross-linking. In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, in particular, Fab fragments, can be accomplished using routine techniques known in the art.

[0466] Chimeric or hybrid antibodies can also be prepared in vitro using known methods of synthetic protein chemistry, including methods involving cross-linking agents. For example, immunotoxins can be constructed using disulfide exchange reactions or by forming thioether bonds. Examples of reagents suitable for this purpose include iminothiol esters and methyl-4-mercaptobutyrimidate. Nucleic acid molecules encoding antibody portions

[0467] In some embodiments, a polynucleotide encoding any of the anti-CD93 constructs or antibody portions described herein is provided. In some embodiments, there is provided a polynucleotide prepared using any of the methods as described herein. In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a heavy or light chain of an antibody portion (eg, an anti-CD93 antibody portion). In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding a heavy chain of an antibody portion (eg, an anti-CD93 antibody portion) and a polynucleotide encoding a light chain of an antibody portion. In some embodiments, the first nucleic acid molecule comprises a first polynucleotide encoding a heavy chain and the second nucleic acid molecule comprises a second polynucleotide encoding a light chain.

[0468] In some such embodiments, the heavy and light chains are represented by one nucleic acid molecule, or as two separate polypeptides, by two separate nucleic acid molecules. In some embodiments, such as when the antibody is a scFv, a single polynucleotide encodes a single polypeptide comprising both the heavy and light chains linked together.

[0469] In some embodiments, a polynucleotide encoding a heavy or light chain of an antibody portion (e.g., an anti-CD93 antibody portion) comprises a nucleotide sequence encoding a leader sequence that is located on the heavy or light chain when translated. N-terminus of the chain. As discussed above, the leader sequence may be a native heavy or light chain leader sequence, or may be another heterologous leader sequence.

[0470] In some embodiments, the polynucleotide is DNA. In some embodiments, the polynucleotide is RNA. In some embodiments, the RNA is mRNA.

[0471] Nucleic acid molecules can be constructed using recombinant DNA techniques well known in the art. In some embodiments, the nucleic acid molecule is an expression vector suitable for expression in the host cell of choice. nucleic acid construct

[0472] In some embodiments, a nucleic acid construct comprising any of the polynucleotides described herein is provided. In some embodiments, a nucleic acid construct prepared using any of the methods described herein is provided.

[0473] In some embodiments, the nucleic acid construct further comprises a promoter operably linked to the polynucleotide. In some embodiments, the polynucleotide corresponds to a gene, wherein the promoter is the wild-type promoter of the gene. carrier

[0474] In some embodiments, a vector is provided comprising any polynucleotide encoding the heavy and / or light chain of any of the antibody portions described herein (e.g., anti-CD93 antibody portions) or Nucleic acid constructs described herein. In some embodiments, a vector prepared using any of the methods described herein is provided. Also provided are vectors comprising a polynucleotide encoding any of the anti-CD93 constructs described herein, such as antibodies, scFv, fusion proteins, or other forms of the constructs (eg, anti-CD93 scFv). Such vectors include, but are not limited to, DNA vectors, phage vectors, viral vectors, retroviral vectors, and the like. In some embodiments, the vector comprises a first polynucleotide sequence encoding a heavy chain and a second polynucleotide sequence encoding a light chain. In some embodiments, the heavy and light chain self-carriers are expressed as two separate polypeptides. In some embodiments, the heavy and light chains are expressed as part of a single polypeptide, such as when the antibody is a scFv.

[0475] In some embodiments, the first vector comprises a polynucleotide encoding the heavy chain and the second vector comprises a polynucleotide encoding the light chain. In some embodiments, the first vector and the second vector are transfected into the host cell in similar amounts (such as similar molar amounts or similar masses). In some embodiments, the first vector and the second vector are transfected into the host cell with a molar or mass ratio between 5:1 and 1:5. In some embodiments, a heavy chain-encoding vector and a light chain-encoding vector are used in a mass ratio between 1:1 and 1:5. In some embodiments, the vector encoding the heavy chain and the vector encoding the light chain are used in a mass ratio of 1:2.

[0476] In some embodiments, a vector is selected that is optimized for expression of the polypeptide in CHO or CHO-derived cells or in NSO cells. Exemplary such vectors are described, eg, in Running Deer et al., Biotechnol. Prog. 20:880-889 (2004). host cell

[0477] In some embodiments, a host cell comprising any of the polypeptides, nucleic acid constructs and / or vectors described herein is provided. In some embodiments, a host cell prepared using any of the methods described herein is provided. In some embodiments, the host cell is capable of producing any of the antibody portions described herein under fermentation conditions.

[0478] In some embodiments, antibody portions described herein (e.g., anti-CD93 antibody portions) can be expressed in prokaryotic cells, such as bacterial cells; or in eukaryotic cells, such as fungal cells (such as yeast), plant cells, Expressed in insect cells and mammalian cells. Such representations can be performed, for example, according to procedures known in the art. Exemplary eukaryotic cells that can be used to express polypeptides include, but are not limited to, COS cells, including COS 7 cells; 293 cells, including 293-6E cells; CHO cells, including CHO-S, DG44. Lec13 CHO cells CHOZN® and FUT8 CHO cells; PER.C6® cells (Crucell); and NSO cells. In some embodiments, antibody portions described herein (eg, anti-CD93 antibody portions) can be expressed in yeast. See, eg, US Publication No. US 2006 / 0270045 Al. In some embodiments, a particular eukaryotic host cell line is selected based on its ability to make desired post-translational modifications to the heavy and / or light chains of the antibody moiety. For example, in some embodiments, a polypeptide produced in CHO cells is more sialylated than the same polypeptide produced in 293 cells.

[0479] Introduction of one or more nucleic acids into a desired host cell can be accomplished by any means including, but not limited to, calcium phosphate transfection, DEAE-polydextrose-mediated transfection, cationic lipid-mediated transfection, Electroporation, transduction, infection, etc. Non-limiting exemplary methods are described, eg, in Sambrook et al., Molecular Cloning, A Laboratory Manual, 3rd Edition Cold Spring Harbor Laboratory Press (2001). Nucleic acids can be transiently or stably transfected into desired host cells according to any suitable method.

[0480] The application also provides host cells comprising any of the polynucleotides or vectors described herein. In some embodiments, the invention provides a host cell comprising an anti-CD93 antibody. Any host cell capable of overexpressing heterologous DNA can be used for the purpose of isolating genes encoding related antibodies, polypeptides or proteins. Non-limiting examples of mammalian host cells include, but are not limited to, COS, HeLa, and CHO cells. See also PCT Publication No. WO 87 / 04462. Suitable non-mammalian host cells include prokaryotes such as E. coli or B. subtilis and yeasts such as S. cerevisae, S. pombe or K. lactis (K. lactis)).

[0481] In some embodiments, antibody portions are produced in a cell-free system. Non-limiting exemplary cell-free systems are described, for example, in Sitaraman et al., Methods Mol. Biol. 498: 229-44 (2009); Spirin, Trends Biotechnol. 22: 538-45 (2004); Endo et al., Biotechnol. Adv. . 21: 695-713 (2003). culture medium

[0482] In some embodiments, a culture medium comprising any of the antibody portions, polynucleotides, nucleic acid constructs, vectors and / or host cells described herein is provided. In some embodiments, a culture medium prepared using any of the methods described herein is provided.

[0483] In some embodiments, the medium comprises hypoxanthine, aminopterin, and / or thymidine (eg, HAT medium). In some embodiments, the medium does not contain serum. In some embodiments, the medium comprises serum. In some embodiments, the medium is D-MEM or RPMI-1640 medium. In some embodiments, the medium is a chemically defined medium. In some embodiments, the chemically defined medium is optimized for the host cell line. Purification of antibody fractions

[0484] Anti-CD93 constructs (eg, anti-CD93 monoclonal antibodies or multispecific antibodies) can be purified by any suitable method. Such methods include, but are not limited to, the use of affinity matrices or hydrophobic interaction chromatography. Suitable affinity ligands include ROR1 ECD and ligands that bind antibody constant regions. For example, protein A, protein G, protein A / G or antibody affinity columns can be used to bind the constant region and purify the anti-CD93 construct comprising the Fc fragment. Hydrophobic interaction chromatography, such as butyl or phenyl columns, may also be suitable for the purification of some polypeptides, such as antibodies. Ion exchange chromatography (eg, anion exchange chromatography and / or cation exchange chromatography) may also be suitable for purifying some polypeptides, such as antibodies. Mixed-mode chromatography (eg, reverse phase / anion exchange, reverse phase / cation exchange, hydrophilic interaction / anion exchange, hydrophilic interaction / cation exchange, etc.) may also be suitable for purification of some polypeptides, such as antibodies. Various methods for purifying polypeptides are known in the art. V. Treatment

[0485] Also provided herein are methods of treating a disease or condition in an individual, or modulating an immune response in an individual. The methods comprise administering an anti-CD93 construct described herein to an individual (eg, a mammal such as a human). It should be understood that the discussion in this section regarding anti-CD93 constructs applies to any anti-CD93 constructs described in this application, such as multispecific anti-CD93 constructs, such as anti-CD93 fusion proteins, such as anti-CD93 / VEGFR fusion proteins , including anti-CD93 / aflibercept fusion protein.

[0486] In some embodiments, provided is a method of treating a disease or condition in an individual, or modulating an immune response in an individual, comprising administering to the individual an effective amount of an anti-CD93 construct described herein. Exemplary diseases or conditions include, but are not limited to, age-related macular degeneration (AMD), diabetic macular edema (DME), choroidal neovascularization (CNV), and cancer.

[0487] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region (VH-2) and a second light chain variable region ( Antibodies or antibody fragments of VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, comprising the amino acid sequence of SEQ ID NO: 2 HC-CDR2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, comprising the LC-CDR1 of SEQ ID NO: 5 LC-CDR2 of the amino acid sequence and LC-CDR3 comprising the amino acid sequence of SEQ ID NO:6. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and iii) comprising the amino acid sequence of SEQ ID NO: 2 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 3, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 4 LC-CDR1 of the amino acid sequence of ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 6, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 2 and iii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 3 CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 4, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 5 and iii) comprising SEQ ID NO: 6 The amino acid sequence of LC-CDR3. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 13, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 14, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0488] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region (VH-2) and a second light chain variable region ( Antibodies or antibody fragments of VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, comprising the amino acid sequence of SEQ ID NO: 18 HC-CDR2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, comprising the LC-CDR1 of SEQ ID NO: 21 LC-CDR2 of the amino acid sequence and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22. In some embodiments, the anti-CD93 VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17 or 304, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 or 305, and iii) HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19, or variants thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and the anti-CD93 VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, 301, 302, 303 or 306, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising SEQ ID NO : LC-CDR3 of an amino acid sequence of 22, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDR.

[0489] In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH Comprising i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18 and iii) comprising the amino acid sequence of SEQ ID NO: 19 HC-CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21 and iii) comprising SEQ ID NO: LC-CDR3 of the amino acid sequence of 22. In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 29 and 307-312, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%). , 96%, 97%, 98% or 99% in any one of the amino acid sequence variants of sequence identity; and VL comprises the amine group of any one of SEQ ID NO: 30 and 313-318 Acid sequence, or comprising amino acids having at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity variants of the sequence.

[0490] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region (VH-2) and a second light chain variable region ( Antibodies or antibody fragments of VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, comprising the amino acid sequence of SEQ ID NO: 34 HC-CDR2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 35, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 36, comprising the LC-CDR1 of SEQ ID NO: 37 LC-CDR2 of the amino acid sequence and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and iii) comprising the amino acid sequence of SEQ ID NO: 34 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of :35, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 36 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 38, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 33, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 34 and iii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 35 CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 36, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 37 and iii) comprising SEQ ID NO: 38 The amino acid sequence of LC-CDR3. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 45, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 46, or comprises at least about 80% (such as at least about 80%, 85% %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0491] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region (VH-2) and a second light chain variable region ( Antibodies or antibody fragments of VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, comprising the amino acid sequence of SEQ ID NO: 66 HC-CDR2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 67, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 68, comprising the LC-CDR1 of SEQ ID NO: 69 LC-CDR2 of the amino acid sequence and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 70. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and iii) comprising SEQ ID NO : HC-CDR3 of the amino acid sequence of 67, or a variant thereof comprising at most 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and the VL comprises i) comprising SEQ ID NO: LC-CDR1 of the amino acid sequence of 68, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 70, or in Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the LC-CDRs. In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 65, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 66 and iii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 67 CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 68, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 69 and iii) comprising SEQ ID NO: 70 The amino acid sequence of LC-CDR3. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 77, or comprises an amino acid sequence having at least about 80% (such as at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%). or 99% of any one) sequence identity variant of the amino acid sequence; and VL comprises the amino acid sequence of SEQ ID NO: 78, or comprises at least about 80% (such as at least about 80%, 85 %, 90%, 95%, 96%, 97%, 98% or 99%) amino acid sequence variants with sequence identity.

[0492] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising an antibody portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the antibody portion is associated with a second heavy chain variable region (VH-2) and a second light chain variable region ( Antibodies or antibody fragments of VL-2) compete for the binding epitope of CD93, wherein the VH-2 comprises HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, comprising the amino acid sequence of SEQ ID NO: 290 HC-CDR2 and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 291, and the VL-2 comprises LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, comprising the LC-CDR1 of SEQ ID NO: 293 LC-CDR2 of the amino acid sequence and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294. In some embodiments, the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and iii) comprising SEQ ID NO HC-CDR3 of the amino acid sequence of : 291, or variants thereof comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the HC-CDR, and VL comprising i) comprising SEQ ID NO: 292 LC-CDR1 of the amino acid sequence of, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 294, or its presence in LC - Variants comprising up to 5, 4, 3, 2 or 1 amino acid substitutions in the CDRs. In some embodiments, the anti-CD93 antibody portion is a humanized antibody derived from an anti-CD93 antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises i) HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 289, ii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290 and iii) HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 291 CDR3, and the VL comprises i) LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, ii) LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and iii) comprising SEQ ID NO: 294 The amino acid sequence of LC-CDR3. In some embodiments, the VH comprises the amino acid sequence of any one of SEQ ID NO: 287 and 319-321, or comprises an amino acid sequence having at least about 80%, such as at least about 80%, 85%, 90%, 95%. , 96%, 97%, 98% or 99% in any one of) the variant of the amino acid sequence of sequence identity; And VL comprises the amine group of any one in SEQ ID NO: 288 and 322-324 Acid sequence, or comprising amino acids having at least about 80% (such as at least about any of 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity variants of the sequence.

[0493] In some embodiments, there is provided a method of treating a disease or condition in an individual, such as AMD, DME, CNV, or cancer, comprising administering to the individual an effective amount of an anti-CD93 construct, the anti-CD93 construct Comprising: a) a full-length antibody that specifically recognizes CD93 comprising two heavy chains and two light chains, wherein each of the two heavy chains comprises a heavy chain variable region (VH), the VH comprising SEQ ID NO: 289 HC-CDR1 of the amino acid sequence, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 290, and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 291, and wherein each of the two light chains comprises a light Chain variable region (VL), this VL comprises the LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 292, the LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 293 and the LC-CD...

Claims

1. An anti-CD93 construct comprising an antibody moiety comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises: HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 17, HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 19; and the VL comprises: LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 20, LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 21, and LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 22, wherein: a) The VH contains the amino acid sequence of SEQ ID NO: 310 or a variant containing an amino acid sequence with at least 80% sequence identity; and the VL contains the amino acid sequence of SEQ ID NO: 318 or a variant containing an amino acid sequence with at least 80% sequence identity; b) The VH contains the amino acid sequence of SEQ ID NO: 29 or a variant containing an amino acid sequence with at least 80% sequence identity; and the VL contains the amino acid sequence of SEQ ID NO: 30 or a variant containing an amino acid sequence with at least 80% sequence identity; c) The VH contains the amino acid sequence of SEQ ID NO: 307 or a variant containing an amino acid sequence with at least 80% sequence identity; and the VL contains the amino acid sequence of SEQ ID NO: 318 or a variant containing an amino acid sequence with at least 80% sequence identity; d) The VH contains SEQ ID NO: e) The VH contains the amino acid sequence of SEQ ID NO: 309 or a variant of the amino acid sequence with at least 80% sequence identity; and the VL contains the amino acid sequence of SEQ ID NO: 318 or a variant of the amino acid sequence with at least 80% sequence identity; or f) The VH contains the amino acid sequence of SEQ ID NO: 312 or a variant of the amino acid sequence with at least 80% sequence identity; and the VL contains the amino acid sequence of SEQ ID NO: 318 or a variant of the amino acid sequence with at least 80% sequence identity.

2. As in Request 1, the anti-CD93 structure, wherein: a) The VH contains the amino acid sequence of SEQ ID NO: 310; and the VL contains the amino acid sequence of SEQ ID NO: 318; b) The VH contains the amino acid sequence of SEQ ID NO: 29; and the VL contains the amino acid sequence of SEQ ID NO: 30; c) The VH contains the amino acid sequence of SEQ ID NO: 307; and the VL contains the amino acid sequence of SEQ ID NO: 318; d) The VH contains the amino acid sequence of SEQ ID NO: 308; and the VL contains the amino acid sequence of SEQ ID NO: 318; e) The VH contains the amino acid sequence of SEQ ID NO: 309; and the VL contains the amino acid sequence of SEQ ID NO: 318; or f) The VH contains the amino acid sequence of SEQ ID NO: 312; and the VL contains the amino acid sequence of SEQ ID NO:

318.

3. The anti-CD93 construct of claim 2, wherein the VH contains the amino acid sequence of SEQ ID NO: 310; and the VL contains the amino acid sequence of SEQ ID NO:

318.

4. An anti-CD93 construct as requested in any of items 1 to 3, wherein the antibody portion is an antibody or its antigen-binding fragment selected from the group consisting of: full-length antibody, bispecific antibody, single-chain Fv (scFv) fragment, Fab fragment, Fab' fragment, F(ab')2, Fv fragment, disulfide bond-stabilized Fv fragment (dsFv), (dsFv)2, Fv-Fc fusion, scFv-Fc fusion, scFv-Fv fusion, bifunctional antibody, trifunctional antibody, and tetrafunctional antibody.

5. An anti-CD93 construct as claimed in any of claims 1 to 3, wherein the antibody portion has an Fc fragment selected from the group consisting of: Fc fragments derived from IgG, IgA, IgD, IgE, IgM, and combinations thereof and mixtures thereof.

6. The anti-CD93 construct as claimed in claim 5, wherein the Fc fragment is selected from the group consisting of Fc fragments derived from IgG1, IgG2, IgG3, IgG4 and combinations thereof and mixtures thereof.

7. The CD93-resistant construct as claimed in claim 5, wherein the Fc fragment has a reduced effect compared to the corresponding wild-type Fc fragment.

8. The CD93-resistant construct as claimed in claim 5, wherein the Fc fragment has an enhanced effect compared to the corresponding wild-type Fc fragment.

9. An anti-CD93 construct as claimed in any of claims 1 to 3, wherein the antibody partially blocks the binding of CD93 to IGFBP7.

10. A CD93-resistant construct as requested in any of items 1 to 3, wherein the CD93 is a human CD93.

11. The anti-CD93 construct of claim 4, wherein the antibody portion comprises: VH comprising the amino acid sequence of SEQ ID NO: 310, and VL comprising the amino acid sequence of SEQ ID NO: 318, wherein the anti-CD93 construct comprises a full-length antibody of the IgG1 Fc fragment.

12. The anti-CD93 construct of claim 4, wherein the antibody portion comprises: a VH comprising the amino acid sequence of SEQ ID NO: 310, and a VL comprising the amino acid sequence of SEQ ID NO: 318, wherein the anti-CD93 construct comprises a bispecific antibody of an IgG1 Fc fragment.

13. A pharmaceutical composition comprising an anti-CD93 construct as claimed in any one of claims 1 to 12 and a pharmaceutically acceptable carrier.

14. An isolated nucleic acid encoding an anti-CD93 construct or its VH or VL as described in any of claims 1 to 12.

15. A vector comprising isolated nucleic acid as claimed in claim 14.

16. An isolated host cell comprising isolated nucleic acid as claimed in claim 14 or a vector as claimed in claim 15.

17. An immune conjugate comprising an antiCD93 construct as claimed in any one of claims 1 to 12, which is attached to a therapeutic agent or a marker.

18. A method for producing an anti-CD93 construct, comprising: a) culturing an isolated host cell as claimed in claim 16 under conditions that effectively express the anti-CD93 construct; and b) obtaining the expressed anti-CD93 construct from the host cell.

19. Use of an anti-CD93 construct as claimed in any one of claims 1 to 12 or a pharmaceutical composition as claimed in claim 13, for the preparation of a pharmaceutical product for treating cancer in an individual.

20. As used in claim 19, wherein the cancer is associated with abnormal vascular structures.

21. As claimed in claim 19, wherein the cancer is a solid tumor or is characterized by tumor hypoxia or is a locally advanced or metastatic cancer.

22. As claimed in claim 19, wherein the cancer comprises a) CD93+ endothelial cells; b) IGFBP7+ blood vessels; and / or c) MMRN2+ blood vessels.

23. As claimed in claim 19, wherein the cancer is selected from the group consisting of: lymphoma, colorectal cancer, brain cancer, breast cancer, ovarian cancer, endometrial cancer, esophageal cancer, prostate cancer, cervical cancer, kidney cancer, bladder cancer, stomach cancer, non-small cell lung cancer, melanoma, and pancreatic cancer.

24. The use as claimed in claim 19, wherein the anti-CD93 construct is administered to the individual not via the intestine, or wherein the use further includes administration of a second therapy.

25. As claimed in claim 24, wherein the second therapy is selected from the group consisting of: surgery, radiation, gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, hormone therapy, targeted therapy, cryotherapy, ultrasound therapy, photodynamic therapy, and chemotherapy.

26. As claimed in claim 25, wherein the immunotherapy comprises administration of an immunomodulatory agent.

27. The use as claimed in claim 26, wherein the immunomodulator is an immune checkpoint inhibitor, and wherein the immune checkpoint inhibitor comprises an anti-PD-L1 antibody or an anti-PD-1 antibody.

28. As used in claim 19, wherein the individual is a human being.

Images