Clinical Research for Azacitidine Combined With Low-dose Dasatinib in Maintenance Therapy of Acute Myeloid Leukemia

Not Applicable

• Conditions: Acute Myeloid Leukemia
• Interventions: Drug: Azacitidine; Drug: Dasatinib

• Registration Number: NCT05042531
• Lead Sponsor: LanZhou University

Brief Summary

This project is a prospective, single-center study to evaluate the efficacy, safety and related mechanisms of azacitidine combined with low-dose dasatinib in maintenance therapy in patients with intermediate and high-risk acute myeloid leukemia(AML). The patients were randomly divided into azacitidine group and azacitidine combined with low-dose dasatinib group. The overall survival and disease-free survival were taken as the main end points, and the mortality and recurrence rate were taken as the secondary end points, meanwhile, the incidence of adverse events were evaluated. At the same time, the mRNA expressions of DNA methyltransferase (DNMT1, DNMT3a, DNMT3b), tumor suppressor genes (TP53, P15, P16, P21, CDH1, DOK6, SHP1, PTPN11) and differentiation genes (pu.1, C/EBP α, C/EBP β) were detected. Pyrophosphate sequencing was used to detect the methylation level of the promoter region of these tumor suppressor genes. Western Blot was used to detect apoptosis proteins (caspase3, caspase8) and phosphorylated proteins (pSTAT3, pSTAT5, pAKT). The proportion of apoptotic population of bone marrow cells was determined by flow cytometry. Therefore, the data in this study will reflect the efficacy and safety of azacitidine or azacitidine combined with low-dose dasatinib in real-world maintenance therapy in patients with medium and high-risk AML.

Detailed Description

In addition to studying the overall survival, disease-free survival and recurrence rates, mortality and incidence of adverse events of patients treated with azacitidine or azacitidine combined with low-dose dasatinib, we will also study its related mechanisms. One of the pathogenesis of AML is that abnormal DNA methylation makes the cell cycle out of control and carcinogenesis by inhibiting the expression of tumor suppressor genes. In addition, the abnormal activation of tyrosine kinase signal pathway also promotes the development of leukemia. Azacitidine, the hypomethylating agents, can not only inhibit the DNA methyltransferase family, but also activate tumor suppressor genes to inhibit a variety of tyrosine kinase signaling pathways, including JAK-STAT. NaShen et al have directly demonstrated that tyrosine kinase inhibitors (TKIs) can not only inhibit the abnormal activation of tyrosine kinase pathway, but also reduce DNA methylation. This study found that the combination of the second generation TKIs and hypomethylating agents can reduce has a synergistic effect on promoting apoptosis and reducing DNA methylation. In addition, TKIs often produces drug resistance due to long exposure time, and the main mechanisms of drug resistance is due to DNA methylation and abnormal reactivation of tyrosine kinase signal pathway. The combination of TKI and azacitidine reduces DNA methylation and inhibits the reactivation of abnormal tyrosine kinase signal pathway, which is helpful to improve TKI drug resistance. Based on the above theory, we assume that patients treated with azacitidine and dasatinib may have more obvious demethylation effect, increased expression of tumor suppressive genes, more obvious apoptosis, and inhibition of phosphorylated protein expression.So we did the lab tests of these mechanisms.We innovatively used azacitidine and TKIs in the treatment of patients with AML maintenance, in order to reduce drug toxicity, enhance drug efficacy, improve patient prognosis and reduce the financial burden of patients.

Study Timeline

• Study First Submitted: 2021-09-01
• Study First Submitted QC: 2021-09-06
• Study First Posted: 2021-09-13
• Study Start: 2021-11-13
• Status Verified: 2022-04
• Last Update Submitted: 2022-04-21
• Last Update Posted: 2022-04-25
• Primary Completion: 2023-09-01
• Study Completion: 2023-12-15

Recruitment & Eligibility

• Status: UNKNOWN
• Sex: All
• Target Recruitment: 30

Inclusion Criteria

1. Patients with intermediate and high-risk AML who are diagnosed according to the 2016 WHO guidelines, aged ≥18 years;

2. Detect minimal residual disease(-) after induction therapy and consolidation therapy;

3. Eastern Cooperative Oncology Group (ECOG) performance status score 0-2;

4. The heart, pulmonary, liver and kidneys have sufficient organ functions:

   1. Cardiac color doppler ultrasound shows cardiac ejection fraction> 50%, heart function classification NYHA III/IV, no heart block or arrhythmia;
   2. Patients without severe restrictive/obstructive pulmonary disease;
   3. Liver function: total bilirubin (TBIL) < 2 times the upper limit of normal, alanine aminotransferase (ALT) and/or aspartate aminotransferase (AST) <2.5 times the upper limit of normal;
   4. Renal function: serum creatinine (Cr) < 1.5 times the upper limit of normal.

5. The patient and family members agree and sign an informed consent form.

Exclusion Criteria

1. Patients with malignant tumors of other organs;

2. HCV positive; or HIV positive; or one of the following HBV test results:

   1. HBsAg positive;
   2. HBsAg negative, HBcAb positive and HBV DNA titer positive;

3. Pregnant and lactating women, and patients who have family planning during the enrollment period;

4. Patients considered to be unsuitable for enrollment by the investigator.

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Arm && Interventions

Group	Intervention	Description
experimental group	Azacitidine	Patients with intermediate and high risk AML were negative for minimal residual disease after intensive induction and consolidation chemotherapy,the patients were randomly divided into two groups, and one group was given azacitidine(75mg/m2, per day on day 1-7\]. Dasatinib 100 mg p.o. qd was administered on days 1-28 of each consolidation cycle.
experimental group	Dasatinib	Patients with intermediate and high risk AML were negative for minimal residual disease after intensive induction and consolidation chemotherapy,the patients were randomly divided into two groups, and one group was given azacitidine(75mg/m2, per day on day 1-7\]. Dasatinib 100 mg p.o. qd was administered on days 1-28 of each consolidation cycle.
control group	Azacitidine	Patients with intermediate and high risk AML were negative for minimal residual disease after intensive induction and consolidation chemotherapy,the patients were randomly divided into two groups, and the other group was given azacitidine(75mg/m2, per day on day 1-7)on days 1-28 of each consolidation cycle.

Primary Outcome Measures

Name	Time	Method
overall survival	up to 30 months.	OS is defined as the time from the date of enrollment until the date of death from any cause.
disease-free survival	up to 30 months.	Event-free survival is defined as the time from enrollment until documented refractory disease, relapse after complete remission(CR) or CR with incomplete recovery of blood counts(CRi), or death from any cause.

Secondary Outcome Measures

Name	Time	Method
methylation level in the promoter region of some tumor suppressor genes.	once before enrollment and once after the completion of the study, up to 30 months.	Detection of methylation level in the promoter region of the above tumor suppressor genes by pyrophosphate sequencing using patient bone marrow aspirate.
mortality	mortality rate at 30 months.	The proportion of patients from enrollment to death was recorded.
recurrence rate	recurrence rate at 30 months.	Record the proportion of patients with recurrence in the study.
adverse events	Adverse events were assessed weekly during the first and second cycles, and every two cycles thereafter (each cycle is 28 days), up to 30 months.	Number of participants with treatment-related adverse events as assessed by CTCAE v5.0.
apoptotic protein and phosphorylated protein	once before enrollment and once after the completion of the study, up to 30 months.	The relative expression of apoptotic protein(caspase3, caspase8) and phosphorylated protein(pSTAT3, pSTAT5, pAKT) were detected by western blot with bone marrow aspirate.
DNA methyltransferase, tumor suppressor genes and differentiation genes	once before enrollment and once after the completion of the study, up to 30 months.	To detect the mRNA expression of DNA methyltransferase(DNMT1, DNMT3a, DNMT3b), tumor suppressor genes(TP53,P15, P16, P21, CDH1, DOK6, SHP1, PTPN11) and differentiation gene(pu.1, C/EBPα, C/EBPβ) with bone marrow aspirate by Q-PCR.
Percentage of bone marrow cell apoptosis population	once before enrollment and once after the completion of the study, up to 30 months.	The percentage of apoptotic population of bone marrow cells was determined by flow cytometry.

Trial Locations

Locations (1)

The First Hospital of Lanzhou University; 🇨🇳 Lanzhou, Gansu, China