Table 1.

Clinical trials with GSI in combination with chemotherapeutic drugs registered at clinicaltrials.gov.

Agent	CT Identifier	Phase	Cancer Type	Drug Combination	Results (as of 1 August 2021)
RO4929097/R4733	NCT01238133	I	Operable Triple-Negative Breast Cancer	Neoadjuvant with Paclitaxel+Carboplatin	n = 14, pCR in 36% of patients, 4 out of 5 patients of higher dose group required dose reduction due to toxicity (neutropenia, thrombocytopenia, hypertension); no paired pre/post-treatment biopsies [391]
	NCT01236586	I	Relapsed/Refractory Solid or CNS tumors, Lymphoma, or T-Cell Leukemia	Dexamethasone	No, withdrawn
	NCT01196416	I/II	Recurrent or Metastatic Melanoma	Cisplatin, Vinblastine, and Temozolomide	n = 14, PR or SD in 8 out of 14 patients which correlated with reduced Notch cleavage in 4 out of 5 analyzed cases of objective response. Adverse effects: leukopenia, thrombocytopenia, elevated transaminases, electrolyte disturbances, hyperglycemia, nausea, vomiting (available at clinicaltrials.gov, accessed on 30 July 2021)
	NCT01119599	I	Malignant Glioma	Temozolomide+radiotherapy	n = 21, MTD was reached (20 mg), no treatment discontinued due to toxicity, generally well-tolerated. PFS 13 months, OS 21 months, better survival correlated with N1IC reduction in post-treatment samples. Reduction of tumor blood perfusion on MRI, significant decrease in N1IC-expressing microvessels without affecting overall microvascular density. The drug had variable BBB penetration with higher concentrations achieved in BBB-disrupted samples. DLL1, DLL3, Jagged2, and HES5 but not HES1 downregulated in post-treatment samples of BBB-disrupted tumors. RO decreased CD133+ CSC pool [390]
	NCT01145456	I	Advanced Solid Tumors	Gemcitabine	n = 18, recommended RO dose for combination with gemcitabine: 30 mg, autoinduction at higher doses, PR in 1 patient (nasopharyngeal carcinoma), SD in 3 patients (pancreas, tracheal, and breast cancer) (n = 18). Adverse effects: elevated transaminases, skin rush, neutropenia. Notch3 levels at IHC were lower in patients who received more than 4 cycles of RO, higher levels of Notch3 in tumor tissue were associated with resistance to RO4929097+gemcitabine [392]
	NCT01158274	I	Refractory Solid Tumors	Capecitabine	n = 30, MTD was not reached, RO autoinduction at high doses, PR in 2 patients (fluoropyrimidine-refractory colon cancer and cervical cancer). Adverse effects: nausea, vomiting, hypophosphatemia, diarrhea [393]
	NCT01192763	I	Pancreatic Cancer	Various neoadjuvant	No, terminated
Nirogacestat/PF-03084014	NCT01876251	I	Advanced Breast Cancer	Docetaxel	n = 29, MTD 100 mg twice daily, PR in 4 and SD in 9 out of 25 patients, median PSF 4.1 months in the expansion cohort. Adverse effects: neutropenia, fatigue, nausea, leukopenia, diarrhea, alopecia, anemia, vomiting. Notch1 and Notch2 RNA in serum decreased on the 2nd day after treatment and increased on the 8th day compared with the baseline. Notch4 RNA in serum decreased on the 8th day [394]
	NCT02109445	I/II	Metastatic Pancreatic Adenocarcinoma	Gemcitabine and Nab-Paclitaxel	n = 3, phase II was not performed, only some pharmacokinetic data posted (available at clinicaltrials.gov, accessed on 31 July 2021)
LY3039478/JSMD194	NCT02784795	I	Advanced or Metastatic Solid Tumors	Cisplatin/Gemcitabine, or Gemcitabine/Carboplatin, or Taladegib, or LY3023414, or Abemaciclib	No
	NCT01695005	I	Advanced or Metastatic Solid Tumors	Prednisone	n = 28, combination aimed to mitigate GSI intestinal toxicity. SD in 54.5% and 64.7% of patients receiving 75 to 150 mg escalating doses of LY TIW (F1) or BIW (F2), respectively. DLT: increased serum amylase, fatigue, hypophosphatemia, maculopapular rush. No DLT in combination with prednisone, GI toxicity less frequent than in no-prednisone groups. In matched pre- and post-treatment tumor samples (n = 10) positive for Notch1 at baseline, 5 were negative for Notch1 post-treatment (2 patients had SD), 2 biopsies remained positive (both SD), and 3 biopsies were not evaluable [395]
	NCT03502577	I	Multiple Myeloma	BCMA-specific CAR T followed with fludarabine and cyclophosphamide	No
	NCT01363817	I	T-ALL or T-LBL	Dexamethasone	No
	NCT02518113	Ib/II	T-ALL/T-LBL	Dexamethasone	n = 36, 1 patient had CR that lasted 10.51 months, 16.7% (n = 6) had SD, 33.3% (n = 12) had PD. 47.2% (n = 17) were not evaluable, median PSF was 1.18 months. MTD: 75 mg LY + 24 mg dexamethasone daily on 1–5 days of treatment. Adverse reactions in 77.8% of patients. Dexamethasone did not revert severe GI adverse events that were registered in 16.7% of patients. DLT: GI hemorrhage, nausea, vomiting, diarrhea. The efficacy of Notch1 cleavage reduction varied from 66% in the group receiving 50 mg of crenigacestat to 87% in the group of 100–125 mg, but higher doses did not correspond to a better clinical outcome [396]
AL101/BMS-906024	NCT01653470	I	Advanced/Metastatic Solid Tumors	Paclitaxel or FOLFIRI or Paclitaxel with and without Carboplatin	No
MK-0752	NCT01098344	I	Inoperable Stage III/IV Pancreatic Cancer	Gemcitabine	n = 44, 13 patients had SD and 1 patient had PR among 19 patients appropriate for tumor response analysis, median time to disease progression was 169 days, median time of overall survival was 246 days. Adverse effects: 55% patients—nausea, 55%—vomiting, 48%—diarrhea, 40.5%—thrombocytopenia, 41%—anemia, 33%—anorexia, 31% -fatigue, 29%—neutropenia. Significant inhibition of Notch signaling in hair follicles was observed in 25/29 patients, no dose-dependent relationship, HES1 expression was evaluated in 20 matched pre/post treatment tumor samples, basal HES1 expression was low, HES1 expression post-treatment was lower in 2 out of 20 biopsy pairs [397]
	NCT00645333	I/II	Advanced/Metastatic Breast Cancer	Docetaxel and Pegfilgrastim	n = 30, of 24 participants evaluable for response, 11 PR, 9 SD, and 3 PD were observed. MTD of MK in combination with docetaxel was 600 mg, 5 cases of DLT, serious adverse effects in 55.3% of patients, adverse effects: 66.67%—fatigue, 50.00%—nausea, 33.33%—diarrhea/hyperglycemia/nail changes, decrease in CD44+/CD24–, ALDH(+) cells in tumors of patients undergoing serial biopsies (3/5) after several cycles of treatment [398]