Novel conjugate molecules targeting cd39 and tgfbeta

The conjugate molecule of any one of preceding claims, wherein the conjugate molecule has one or more properties selected from the group consisting of:

• a) specifically binding to human CD39 but not specifically binding to mouse CD39 as measured by FACS assay;
• b) specifically binding to cynomolgus CD39 at an EC50 of no more than 10−8 M as measured by FACS assay;
• c) specifically binding to human CD39 at a KD value of no more than 10−7 M (e.g. no more than 5×10−8 M, no more than 3×10−8 M, no more than 2×10−8 M, no more than 1×10−8 M, or no more than 8×10−9 M) as measured by Biacore assay;
• d) specifically binding to human CD39 at a KD value of no more than 10−8 M (e.g. no more than 8×10−9 M, no more than 5×10−9 M, no more than 4×10−9 M, no more than 3×10−9 M, no more than 1×10−9 M, or no more than 9×10−10 M) as measured by Octet assay;
• e) inhibiting ATPase activity in a CD39 expressing cell at an IC 50 of no more than nM (e.g. no more than 1 nM, no more than 5 nM, no more than 10 nM, or no more than 30 nM) as measured by ATPase activity assay;
• f) capable of enhancing ATP mediated monocytes activation at a concentration of no more than 10 nM (e.g. no more than 5 nM, no more than 3 nM, no more than 2 nM, no more than 1 nM, no more than 0.5 nM, or no more than 0.2 nM) as measured by analysis of CD80, CD86 and/or CD40 expression by FACS assay;
• g) capable of enhancing ATP mediated T cell activation in PBMC at a concentration of no more than 25 nM as measured by IL-2 secretion, IFN-γ secretion, CD4+ or CD8+ T cells proliferation;
• h) capable of enhancing ATP mediated dendritic cell (DC) activation at a concentration of no more than 25 nM (or no more than 10 nM, or no more than or no more than 1 nM, or no more than 0.5 nM) as measured by analysis of CD83 expression by FACS assay, or by the capability of the activated DC to promote T cell proliferation, or by the capability of the activated DC to promote IFN-γ production in the mix-lymphocyte reaction (MLR) assay;
• i) capable of blocking the inhibition of CD4+ T cell proliferation induced by adenosine (hydrolyzed from ATP) at a concentration of no more than 1 nM (e.g. no more than 0.1 nM, no more than 0.01 nM) as measured by FACS assay;
• j) capable of inhibiting tumor growth in a mammal a NK cell or macrophage cell dependent manner;
• k) capable of reversing human CD8+ T cell proliferation which was inhibited by eATP as measured by T cell proliferation, CD25+ cells, and living cells population;
• and
• l) capable of enhancing human macrophage IL1β release induced by LPS stimulation at a concentration of no more than 50 nM (or no more than 12.5 nM, or no more than 3.13 nM, or no more than 0.78 nM, or no more than 0.2 nM, or no more than 0.049 nM, or no more than 0.012 nM, or no more than 0.003 nM, or no more than 0.0008 nM) as measured by ELISA assay.

The conjugate molecule of any one of the preceding claims, wherein the conjugate molecule has one or more properties selected from the group consisting of:

• a) specifically binding to human TGFβ1, human TGFβ2, and/or human TGFβ3;
• b) specifically binding to human TGFβ1 and mouse TGFβ1 with similar affinity;
• c) specifically binding to human TGFβ1 at an EC50 of no more than 3×10−11 M (e.g. no more than 2×10−11 M, no more than 1×10−11 M, no more than 0.9×10−11 M, no more than 0.8×10−11 M, no more than 0.7×10−11 M, no more than 0.6×10−11 M, no more than 0.5×10−11 M) as measured by ELISA assay;
• d) capable of blocking human TGFβ1 and TGFβRII binding at an IC50 of no more than 4×10−10 M (e.g. no more than 3×10−10 M, no more than 2×10−10 M, no more than 1×10−10 M, no more than 0.5×10−10 M) as measured by blocking assay;
• e) capable of binding to human CD39 in a dose-dependent manner as measured by FACS assay;
• f) capable of simultaneously binding to CD39 and TGFβ as measured by ELISA assay or FACS assay;
• g) capable of inhibiting TGFβ signal at an IC50 no more than 4×10−11 M as measured by a TGF-β SMAD reporter assay;
• h) capable of inhibiting ATPase activity in a CD39 expressing cell at an IC50 of no more than 7×10−10 m (e.g. no more than 6×10−10 M, no more than 5×10−10 M, no more than 4×10−10 M, no more than 3×10−10 M, no more than 2×10−10 M, no more than 1×10−10 M, no more than 0.5×10−10 M) as measured by ATPase activity assay;
• i) specifically binding to human CD39 at a KD value of no more than 4×10−10 M (e.g. no more than 3×10−10 M, no more than 2×10−10 M, no more than 1×10−10 M, or no more than 0.5×10−10 M) as measured by Octet assay;
• j) specifically binding to human TGFβ1 at a KD value of no more than 4×10−11 M (e.g. no more than 3×10−11 M, no more than 2×10−11 M, no more than 1×10−11 M, or no more than 0.5×10−11 M) as measured by Octet assay;
• k) capable of recovering T cell function as measured by a Treg suppression assay;
• l) capable of inhibiting human T cell apoptosis in a dose-dependent way;
• m) capable of promoting human T cell survival and activation over stimulation;
• n) capable of blocking TGFβ induced Foxp3 expression on total T cells;
• and
• o) capable of restoring ATP induced inhibition on human T cell proliferation.