Combination of venetoclax and azacitidine in relapsed/refractory acute B-cell lymphoblastic leukemia: a case series from a single center.

OBJECTIVES: Relapsed/refractory acute B-cell lymphoblastic leukemia (R/R B-ALL) often responds poorly to induction chemotherapy. However, recent research has shown a novel and effective drug treatment for R/R B-ALL. METHODS: A total of eight patients with R/R B-ALL were enrolled in the study from November 2021 to August 2022. All patients received chemotherapy based on a combination regimen of venetoclax and azacitidine. The regimen was as follows venetoclax 100 mg d1, 200 mg d2, 400 mg d3-14, azacitidine 75 mg/m2 d1-7. RESULTS: Five of eight patients achieved very deep and complete remission (CR) with minimal residual disease (MRD) less than 0.1%. One patient achieved partial remission. Two patients did not achieve remission. There were no serious adverse events and all patients were well tolerated. Three patients were eligible for consolidation chemotherapy and were bridged to CAR-T therapy. CONCLUSIONS: The combined regimen of venetoclax and azacitidine may be beneficial for patients with R/R B-ALL.

Hsa-circ_0003420 induces apoptosis in acute myeloid leukemia stem cells and impairs stem cell properties.

OBJECTIVE: To explore the effect of circular RNA-0003420 (circ_0003420) in acute myeloid leukemia (AML) relapse and leukemia stem cells (LSCs) properties. MATERIALS AND METHODS: TRIzol reagent was used to extract total RNA from AML tissues or cells. Cell viability was assessed by Cell Counting Kit-8 and EdU staining assays. Cell apoptosis was determined by flow cytometry. RESULTS: Compared with normal hematopoietic stem cells, circular RNA hsa-circ_0003420 expression was considerably decreased in non-m3 AML stem cells. Furthermore, the lack of hsa-circ_0003420 is correlated with poor clinical results and impaired therapeutic effects in AML. Overexpression of hsa-circ_0003420 via transfection caused LSC death and inhibited the characteristics of leukemia tumor stem cells, including expression of Homeobox B4 (HOXB4), MYB, and aldehyde dehydrogenase 1 family member A1 (ALDH1A1) axis. Furthermore, hsa-circ_0003420 targets the mRNA of insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) and hsa-circ_0003420 expression markedly repressed IGF2BP1 levels in LSCs. Restoration of IGF2BP1 eliminated the effect of hsa-circ_0003420 on the replication, apoptosis, and LSC phenotype of KG-1a cells. DISCUSSION AND CONCLUSIONS: Up-regulation of hsa-circ_0003420 expression in LSCs caused redox disorder, inflammation and apoptosis, suggesting that this protein could be used as a target for the treatment AML.

Association between miR-212-3p and SOX11, and the effects of miR-212-3p on cell proliferation and migration in mantle cell lymphoma.

To the best of our knowledge, the effect of miR-212-3p on sex-determining region Y-box 11 (SOX11) expression has not been previously investigated and how this effect affects cell proliferation and migration in lymphoma remains unclear. The present study aimed to assess the association between microRNA-212-3p (miR-212-3p) and SOX11, and the effects of miR-212-3p on cell proliferation and migration in mantle cell lymphoma. Cancer tissue and corresponding paracancerous tissue samples were collected from 65 patients with mantle cell lymphoma. The mRNA expression levels of miR-212-3p and SOX11 were analyzed using quantitative PCR, and SOX11 protein expression was determined using western blotting. Following transfection, the miR-212-3p mimic group exhibited a significantly lower SOX11 mRNA and protein expression than the miR-NC group. After 48-72 h of transfection, cell proliferation in the miR-212-3p mimic group was significantly lower than that in the miR-NC group. Furthermore, the miR-212-3p mimic group exhibited significantly lower cell invasion and significantly higher apoptosis than the miR-NC group. The current results suggested that miR-212-3p inhibited lymphoma cell proliferation and migration, and promoted their apoptosis by specifically regulating SOX11. Therefore, miR-212-3p may serve as a novel therapeutic target and marker for lymphoma.

Two new phenylethanoid glycosides from Ginkgo biloba leaves and their tyrosinase inhibitory activities.

Two undescribed phenylethanoid glycosides, Ginkgoside C (1) and D (2), together with ten known glycosides (3-12) were isolated from Ginkgo biloba leaves. Their structures were characterized by physical data analyses such as NMR, HRESIMS, as well as chemical hydrolysis. All compounds were tested for their tyrosinase inhibitory activities. At a concentration of 25 µM, compounds 2, 4, 5, 6, and 11 showed obvious mushroom tyrosinase inhibition activities, with %inhibition values of 19.12 ± 2.59%, 25.79 ± 1.83%, 16.07 ± 1.07%, 24.46 ± 1.10%, 18.64 ± 3.62%, respectively, with kojic acid used as the positive control (27.50 ± 2.72%).

Isolation, structure elucidation, tyrosinase inhibitory, and antioxidant evaluation of the constituents from Angelica dahurica roots.

Two undescribed phenolic compounds, angelicols A (1) and B (2) and one undescribed coumarin rhamnoside, angelicoside A (3), together with 17 known compounds (4-20) were isolated from the roots of Angelica dahurica. Their structures were characterized by physical data analyses such as NMR, HRESIMS, and X-ray diffraction. Compounds 2, 3, 5, 6 and L-ascorbic acid (positive control) exhibited obvious DPPH radical scavenging activities with IC50 values of 0.36 mM, 0.43 mM, 0.39 mM, 0.44 mM, 0.25 mM, respectively. At a concentration of 25 µM, all compounds showed weaker tyrosinase inhibition activities (%inhibition < 5%) than kojic acid (26.00 ± 0.67%, IC50 = 44.29 ± 0.06 µM).

The chronic administration of two novel long-acting Xenopus glucagon-like peptide-1 analogs xGLP159 and xGLP296 potently improved systemic metabolism and glycemic control in rodent models.

We here reported 2 novel Xenopus glucagon-like peptide-1 (xGLP-1) analogs, xGLP159 and xGLP296, whose therapeutic effects on metabolic efficacy and glycemic control were evaluated in rodents. The in vitro potency of xGLP159 and xGLP296 were investigated on human embryonic kidney 293 cells. The acute glucose-lowering and insulinotropic effects of xGLP159 and xGLP296 were assessed in the Institute of Cancer Research, Kunming, and diabetic (db/db) mice. The pharmacokinetic profiles of xGLP159 and xGLP296 were confirmed on Sprague-Dawley (SD) rats and their long-acting hypoglycemic and anorectic effects were evaluated in db/db mice. Chronic treatment effects of xGLP159 and xGLP296 were evaluated in diet-induced obese (DIO) mice and db/db mice. The results showed that xGLP159 and xGLP296 exhibited comparable receptor activation potency, hypoglycemic effect, and insulinotropic activity to liraglutide. The enhanced half-lives of xGLP159 and xGLP296 in SD rats (5.1 and 5.8 h, respectively) resulted in prolonged anti-db/db durations in db/db mice. Three weeks' administration of xGLP159 and xGLP296 normalized glucose tolerance and adiposity in DIO mice. Furthermore, 11-wk treatment of xGLP159 and xGLP296 corrected hyperglycemia and improved pancreatic function in db/db mice. These preclinical studies supported xGLP159 and xGLP296 as promising candidates for the treatment of metabolic diseases.-Han, J., Meng, T., Chen, X., Han, Y., Fu, J., Zhou, F., Fei, Y., Li, C. The chronic administration of two novel long-acting Xenopus glucagon-like peptide-1 analogs xGLP159 and xGLP296 potently improved systemic metabolism and glycemic control in rodent models.

Rational design of dimeric lipidated Xenopus glucagon-like peptide 1 analogues as long-acting antihyperglycaemic agents.

Dimerization is viewed as an effective means to enhance the binding affinity and therapeutic potency of peptides. Both dimerization and lipidation effectively prolong the half-life of peptides in vivo by increasing hydrodynamic size and facilitating physical interactions with serum albumin. Here, we report a novel method to discover long-acting glucagon-like peptide 1 (GLP-1) analogues by rational design based on Xenopus GLP-1 through a combined dimerization and lipidization strategy. On the basis of our previous structure analysis of Xenopus GLP-1, palmitic acid and a C-terminal Cys were firstly introduced into two Xenopus GLP-1 analogues (1 and 2), and the afforded 3 and 4 were further reacted with bis-maleimide amine to afford two dimeric lipidated Xenopus GLP-1 analogues (5 and 6). The in vitro and in vivo biological activities of 5 and 6 were significantly improved as compared with their monomers. Moreover, the selected compound 6 showed greater hypoglycemic and insulinotropic activities than liraglutide even when the dose of 6 was reduced to half in db/db mice. Pharmacokinetic test revealed that 6 had a ∼ 3-fold longer half-life than liraglutide in Kunming mice and SD rats, and the longer half-life of 6 led to excellent long-acting hypoglycemic effects as confirmed by two different pharmacological methods conducted on db/db mice. Finally, a 7 weeks chronic study conducted on db/db mice demonstrated the better therapeutic efficacies of 6 on glucose tolerance normalization, HbA1c reduction and pancreas islets protection than liraglutide. The present research showed that combined dimerization and lipidization is effective when applied to Xenopus GLP-1 analogue to develop novel GLP-1 analogue for the treatment of type 2 diabetes. In addition, the promising preclinical data of 6 suggested the therapeutic potential of 6 as a novel anti-diabetic agent.

Lipidation and conformational constraining for prolonging the effects of peptides: Xenopus glucagon-like peptide 1 analogues with potent and long-acting hypoglycemic activity.

The main disadvantages of glucagon-like peptide 1 (GLP-1) are its rapid degradation and excretion. These bottlenecks can be overcome by lipidation or other structural modification. The aim of this study was to design a series of long-acting GLP-1 analogues based on our previously discovered Xenopus GLP-1 analogs (1-3). The structure-activity relationship around lipidated 1-3 derivatives (1a-3l) with respect to in vitro potency as well as protraction was firstly explored. Compound 3g was selected for further modification. The Gly2 of 3g was replaced with Aib2, and a lactam constraint between Glu16 and Lys20 (i to i + 4) was introduced to further improve the in vivo activity and stability, affording compound 4. The receptor activation capability and in vitro stability of 4 were better than 3g and liraglutide. In addition, the hypoglycemic and insulinotropic activity of 4 was significantly better than liraglutide in db/db mice. Moreover, the enhanced in vitro stability of 4 translated into improved in vivo pharmacokinetic profiles and a prolonged antidiabetic duration. Administration of 4 twice daily for one week in diet-induced obese mice caused a significant decrease in food intake, body fat and body weight. The five-week treatment study on db/db mice of 4 further demonstrated the therapeutic effects of 4 on body weight, HbA1c and glucose tolerance. These preclinical studies demonstrate the therapeutic potential of 4 for type 2 diabetes and obesity. The present study also suggests that combined lipidation and conformational constraint strategy has potential to be used for improving the therapeutic properties of peptides.

Lithocholic Acid-Based Peptide Delivery System for an Enhanced Pharmacological and Pharmacokinetic Profile of Xenopus GLP-1 Analogs.

GLP-1 analogs suffer from the main disadvantage of a short in vivo half-life. Lithocholic acid (LCA), one of the four main bile acids in the human body, possesses a high albumin binding rate. We therefore envisioned that a LCA-based peptide delivery system could extend the half-life of GLP-1 analogs by facilitating the noncovalent binding of peptides to human serum albumin. On the basis of our previously identified Xenopus GLP-1 analogs (1-3), a series of LCA-modified Xenopus GLP-1 conjugates were designed (4a-4r), and the bioactivity studies of these conjugates were performed to identify compounds with balanced in vitro receptor activation potency and plasma stability. 4c, 4i, and 4r were selected, and their LCA side chains were optimized to further increase their stability, affording 5a-5c. Compound 5b showed a more increased albumin affinity and prolonged in vitro stability than that of 4i and liraglutide. In db/ db mice, 5b exhibited comparable hypoglycemic and insulinotropic activity to liraglutide and semaglutide. Importantly, the enhanced albumin affinity of 5b resulted in a prolonged in vivo antidiabetic duration. Finally, chronic treatment investigations of 5b demonstrated the therapeutic effects of 5b on HbA1c, body weight, blood glucose, and pancreatic endocrine deficiencies on db/ db mice. Our studies revealed 5b as a promising antidiabetic candidate. Furthermore, our study suggests the derivatization of Xenopus GLP-1 analogs with LCA represents an effective strategy to develop potent long-acting GLP-1 receptor agonists for the treatment of type 2 diabetes.

Preparation and Pharmaceutical Characterizations of Lipidated Dimeric Xenopus Glucagon-Like Peptide-1 Conjugates.

A pair of glucagon-like peptide-1 (GLP-1) analogs (1 and 2) were synthesized by hybridizing the key sequences of GLP-1, exendin-4, lixisenatide, and xenGLP-1B (Xenopus GLP-1 analog). To achieve long-acting hypoglycemic effects and to further improve their anti-diabetic potencies, lipidization and dimerization strategies were used to afford two lipidated dimeric conjugates (9 and 11). Conjugates 9 and 11 showed stronger receptor activation potency than GLP-1 and exendin-4 in vitro. Moreover, 9 and 11 exhibited superior hypoglycemic and insulinotropic activities to liraglutide in type 2 diabetic C57BL/6J-m+/+ Leprdb (db/db) mice. Pharmacokinetic studies revealed that the circulating half-lives (t1/2) of 9 and 11 were 2.3- and 1.7-fold longer than that of liraglutide. The improved pharmacokinetic profiles led to significantly protracted in vivo anti-diabetic effects as confirmed by multiple oral glucose tolerance tests and hypoglycemic duration tests. Most importantly, chronic treatment studies found that once daily administration of 9 or 11 in db/db mice achieved more beneficial effects on HbA1c reduction and glucose tolerance normalization than liraglutide. Our research demonstrated lipidization and dimerization as useful tools for the development of novel GLP-1 receptor agonists. The preclinical studies suggested the potential of 9 and 11 to be developed as novel anti-diabetic agents.

Xenopus-derived glucagon-like peptide-1 and polyethylene-glycosylated glucagon-like peptide-1 receptor agonists: long-acting hypoglycaemic and insulinotropic activities with potential therapeutic utilities.

BACKGROUND AND PURPOSE: Incretin-based therapies based on glucagon-like peptide-1 (GLP-1) receptor agonists are effective treatments of type 2 diabetes. Abundant research has focused on the development of long-acting GLP-1 receptor agonists. However, all GLP-1 receptor agonists in clinical use or development are based on human or Gila GLP-1. We have identified a potent GLP-1 receptor agonist, xGLP-1B, based on Xenopus GLP-1. EXPERIMENTAL APPROACH: To further modify the structure of xGLP-1B, alanine scanning was performed to study the structure -activity relationship of xGLP-1B. Two strategies were then employed to improve bioactivity. First, the C-terminal tail of lixisenatide was appended to cysteine-altered xGLP-1B analogues. Second, polyethylene glycol (PEG) chains with different molecular weights were conjugated with the peptides, giving a series of PEGylated conjugates. Comprehensive bioactivity studies of these conjugates were performed in vitro and in vivo. RESULTS: From the in vitro receptor activation potency and in vivo acute hypoglycaemic activities of conjugates 25 -36, 33 was identified as the best candidate for further biological assessments. Conjugate 33 exhibited prominent hypoglycaemic and insulinotropic activities, as well as improved pharmacokinetic profiles in vivo. The prolonged antidiabetic duration of 33 was further confirmed by pre-oral glucose tolerance tests (OGTT) and multiple OGTT. Furthermore, chronic treatment of db/db mice with 33 ameliorated non-fasting blood glucose and insulin levels, reduced HbA1c values and normalized their impaired glucose tolerance. Importantly, no in vivo toxicity was observed in mice treated with 33. CONCLUSIONS AND IMPLICATIONS: Peptide 33 is a promising long-acting type 2 diabetes therapeutic deserving further investigation.

Micellar Nanomedicine of Novel Fatty Acid Modified Xenopus Glucagon-like Peptide-1: Improved Physicochemical Characteristics and Therapeutic Utilities for Type 2 Diabetes.

To develop novel long-acting antidiabetics with improved therapeutic efficacy, two glucagon-like peptide-1 (GLP-1) analogs were constructed through the hybridization of key sequences of GLP-1, xenGLP-1B, exendin-4, and lixisenatide. Hybrids 1 and 2 demonstrated enhanced in vitro and in vivo biological activities and were further site-specifically lipidized at lysine residues to achieve prolonged duration of action and less frequent administration. Compared with their native peptides, compounds 3-6 showed similar in vitro activities but impaired in vivo acute hypoglycemic potencies due to decreased aqueous solubility and retarded absorption in vivo. To circumvent these issues, compound 3 (xenoglutide) was selected to be self-associated with sterically stabilized micelles (SSM). The α-helix and solubility of xenoglutide were significantly improved after self-associated with SSM. Notably, the improved physicochemical characteristics of xenoglutide-SSM led to revival of acute hypoglycemic ability without affecting its long-term glucose-lowering activity. Most importantly, preclinical studies demonstrated improved therapeutic effects and safety of xenoglutide-SSM in diabetic db/db mice. Our work suggests the SSM incorporation as an effective approach to improve the pharmacokinetic and biological properties of hydrophobicity peptide drugs. Furthermore, our data clearly indicate xenoglutide-SSM as a novel nanomedicine for the treatment of type 2 diabetics.

Xenopus GLP-1-inspired discovery of novel GLP-1 receptor agonists as long-acting hypoglycemic and insulinotropic agents with significant therapeutic potential.

We here report the discovery and therapeutic efficacy of a novel series of glucagon-like peptide-1 (GLP-1) receptor agonists derived from Xenopus GLP-1. First, five amino acid-mutated Xenopus GLP-1s were synthesized, and xGLP-3 with the best acute and long-acting hypoglycemic activity was selected for further modification. Next, PEGylation of xGLP-3 was performed at specific sites, which were determined using cysteine mutagenesis scanning. Twelve PEGylated conjugates tethered with Mal-PEGs of 1, 2, and 5kDa were synthesized. Conjugates 7b and 7c, which exhibited comparable hypoglycemic and insulinotropic effects to Gly8-GLP-1, were selected for in-depth evaluation. It was found that 7b and 7c exhibited prolonged in vivo half-life and improved pharmacokinetic behaviors. The long-term hypoglycemic effects of 7b and 7c were further confirmed by pre-OGTT and multiple OGTT. Importantly, long-term administration of 7b or 7c in db/db mice achieved beneficial effects on body weight loss, food intake and HbA1c reduction, and glucose tolerance normalization. These preclinical studies indicate the promising role of 7b and 7c as long-acting type 2 diabetes therapeutics. In addition, our research demonstrated the feasibility of developing novel antidiabetic agents based on Xenopus GLP-1.

Design, synthesis and biological evaluation of PEGylated Xenopus glucagon-like peptide-1 derivatives as long-acting hypoglycemic agents.

In order to develop novel long-acting GLP-1 derivatives, a peptide hybrid (1a) from human GLP-1 and Xenopus GLP-1 discovered in our previous research was selected as the lead compound. Exendin-4 inspired modification resulted in peptide 1b with enhanced glucose-lowering activity. Cysteine mutated 1b derivatives with reserved bioactivity were further site-specifically connected with mPEG2000-MAL to provide conjugates 3a-h, among which 3d and 3e were found to have significantly improved hypoglycemic activity and insulinotropic ability than GLP-1. The hypoglycemic durations of 3d and 3e were remarkably prolonged to ∼20 h in type 2 diabetic db/db mice, compared with the 5.3 h of exendin-4 in the same test. Finally, chronic in vivo studies revealed that a once-daily treatment of 3d or 3e for five weeks resulted in recovered glucose-controlling ability of type 2 diabetic db/db mice, along with other benefits, such as reduced body weight gains, food intake amounts and HbA1c values. Collectively, our results suggest 3d and 3e as potential long-acting glucose-lowering agents for treating type 2 diabetes.