Beijng Immunoah Pharma Tech Co., Ltd

Vulvar mesotherapy enables targeted drug delivery to the reproductive tract through the "first-pass effect". We administered human long-acting recombinant follicle-stimulating hormone (LArF) via vulvar mesotherapy (LArF-VM) for ovarian stimulation in pre-synchronized Holstein cows. The single-dose LArF-VM protocol was compared with the conventional intramuscular multi-injection short-acting porcine pituitary FSH (SApF-IM) protocol. The objectives were to optimize the LArF-VM dose, validate the optimal dose against the conventional protocol, and evaluate operational efficiency, costs, and welfare benefits. In the dose-optimisation experiment (n = 20), 150 µg LArF-VM gave significantly greater total follicles (18.6 ± 3.84), medium-sized follicles (12.8 ± 2.78), and retrieved oocytes (15.6 ± 2.70) compared to 100 µg or the SApF-IM protocol (P ≤ 0.05). The 150 µg dose outperformed all groups in oocyte recovery and produced a favourable follicle distribution (21.74% small, 69.57% medium, 8.70% large) versus SApF-IM (10.77% small, 49.23% medium, 40% large) (P ≤ 0.05). The LArF-VM protocol significantly reduced the number of handling rounds, injection time, diluent volume, and labour cost per donor cow (P ≤ 0.01). Validation experiment (n = 13) showed that follicular response (22.17 ± 7.9 vs 21.71 ± 9.4) and oocyte yield (15.67 ± 6.02 vs 15.86 ± 7.15) did not differ significantly between the LArF-VM and SApF-IM protocols (P > 0.05). The single-injection LArF-VM protocol offers a simpler, safer, less stressful, logistically superior, and cost-effective alternative for ovum pick-up while maintaining effective follicular stimulation and oocyte recovery in cattle.

Colorectal cancer is a major global health burden, with limited efficacy of traditional treatment modalities in improving survival rates. However, recently advances in immunotherapy has improved treatment outcomes for patients with this cancer. To address the continuing need for improved treatment efficacy, this study introduced a novel tri-specific antibody, IMT030122, that targets EpCAM, 4-1BB, and CD3. We evaluated the pharmacological efficacy and mechanism of action of IMT030122 in vitro and in vivo. In in vitro studies, IMT030122 exhibited differential binding to antigens and cells expressing EpCAM, 4-1BB, and CD3. Moreover, IMT030122 relied on EpCAM-targeted activation of intracellular CD3 and 4-1BB signaling and mediated T cell cytotoxicity specific to HCT116 colorectal cancer cells. In vivo, IMT030122 demonstrated potent anti-tumor activity, significantly inhibiting the growth of colon cancer HCT116 and MC38-hEpCAM subcutaneous grafts. Further pharmacological analysis revealed that IMT030122 recruited lymphocytes from peripheral blood into colorectal cancer tissue and exerted durable anti-tumor activity, predominantly by promoting the activation, proliferation, and differentiation of CD8T cells. Notably, IMT030122 still exhibited anti-tumor efficacy even in the presence of significantly depleted lymphocytes in colorectal cancer tissue. The potent pharmacological activity and anti-tumor effects of IMT030122 suggest it may enhance treatment efficacy and substantially extend the survival of patients with colorectal cancer in the future.

Additional immunotherapies are still warranted for non-responders to checkpoint inhibitors with refractory or relapsing cancers, especially for patients with "cold" tumours lacking significant immune infiltration at treatment onset. We developed XFab-α4-1BB/CD40L, a bispecific antibody targeting 4-1BB and CD40 for dendritic cell activation and priming of tumour-reactive T cells to inhibit tumours.

XFab-α4-1BB/CD40L was developed by engineering an anti-4-1BB Fab arm into a CD40L trimer based on XFab® platform. Characterisation of the bispecific antibody was performed by cell-based reporter assays, maturation of dendritic cell assays, and mixed lymphocyte reactions. The abilities of antigen-specific T-cell expansion and antitumour efficacy were assessed in syngeneic mouse tumour models. Toxicological and pharmacodynamic profiles were investigated in non-human primates.

XFab-α4-1BB/CD40L demonstrated independent CD40 agonistic activity and conditional 4-1BB activity mediated by CD40 crosslinking, leading to dendritic cell maturation and T-cell proliferation in vitro. We confirmed the expansion of antigen-specific T cells in the vaccination model and potent tumour regression induced by the bispecific antibody alone or in combination with gemcitabine in vivo, concomitant with improved tumour-reactive T-cell infiltration. XFab-α4-1BB/CD40L showed no signs of liver toxicity at doses up to 51 mg/kg in a repeated-dose regimen in non-human primates.

XFab-α4-1BB/CD40L is capable of enhancing antitumour immunity by modulating dendritic cell and T-cell functions via targeting 4-1BB agonism to areas of CD40 expression. The focused, potent, and safe immune response induced by the bispecific antibody supports further clinical investigations for the treatment of solid tumours.