Systematic Chemical Mutagenesis Identifies a Potent Novel Apratoxin A/E Hybrid with Improved in Vivo Antitumor Activity

Apratoxins are cytotoxic marine natural products that prevent cotranslational translocation early in the secretory pathway. We showed that apratoxins downregulate receptors and growth factor ligands, giving a one–two punch to cancer cells, particularly those that rely on autocrine loops. Through total synthesis, we tested the effects of amino acid substitutions, including alanine scanning, on the downregulation of receptor tyrosine kinases and vascular endothelial growth factor A (VEGF-A) and probed the stereospecificity of target engagement by epimerization of selected chiral centers. Differential effects on two types of secretory molecules suggest that the apratoxins' substrate selectivity with respect to inhibition of secretion may be tuned through structural modifications to provide tailored therapy. Our structure–activity relationship studies and medicinal chemistry efforts led to a potent inhibitor with in vivo efficacy in a colorectal tumor xenograft model without irreversible toxicity exerted by apratoxin A, demonstrating that this novel mechanism of action has therapeutic potential

Improved Total Synthesis and Biological Evaluation of Potent Apratoxin S4 Based Anticancer Agents with Differential Stability and Further Enhanced Activity

Apratoxins are cytotoxic natural products originally isolated from marine cyanobacteria that act by preventing cotranslational translocation early in the secretory pathway to downregulate receptor levels and inhibit growth factor secretion, leading to potent antiproliferative activity. Through rational design and total synthesis of an apratoxin A/E hybrid, apratoxin S4 (<b>1a</b>), we have previously improved the antitumor activity and tolerability in vivo. Compound <b>1a</b> and newly designed analogues apratoxins S7–S9 (<b>1b</b>–<b>d</b>), with various degrees of methylation at C34 (<b>1b</b>,<b>c</b>) or epimeric configuration at C30 (<b>1d</b>), were efficiently synthesized utilizing improved procedures. Optimizations have been applied to the synthesis of key intermediate aldehyde <b>7</b> and further include the application of Leighton’s silanes and modifications of Kelly’s methods to induce thiazoline ring formation in other crucial steps of the apratoxin synthesis. Apratoxin S9 (<b>1d</b>) exhibited increased activity with subnanomolar potency. Apratoxin S8 (<b>1c</b>) lacks the propensity to be deactivated by dehydration and showed efficacy in a human HCT116 xenograft mouse model

Table1_Characteristics and treatment strategies of aggressive angiomyxoma in women: A retrospective review of 87 cases.docx

ObjectiveAggressive angiomyxoma (AAM) is a rare kind of soft tissue tumor. The purpose of this study is to summarize the clinical manifestations and treatment strategy of AAM in women.MethodWe searched the case reports on AAM in EMBASE, Web of Science and PubMed, China biomedical database, Wanfang database, VIP database, and China National Knowledge Internet from the start of database construction to November 2022 without any language restrictions in place. Then, the obtained case data were extracted, summarized, and analyzed.ResultA total of 74 articles were retrieved involving 87 cases. The age ranges of onset were 2–67 years. The median age at onset was 34 years. The size of the tumor varied greatly among individuals, and about 65.5% of them were asymptomatic. MRI, ultrasound, and needle biopsy were used for diagnosis. Surgery was the primary mode of treatment, but it was prone to relapse. Gonadotropin-releasing hormone agonist (GnRH-a) might be used to reduce the tumor size before the operation and prevent recurrence after the operation. For patients who are unwilling to receive surgical treatment, GnRH-a alone could be attempted.ConclusionDoctors should consider the possibility of AAM in women with genital tumors. For surgery, it must be ensured that the negative surgical margin is recommended and achieved for preventing recurrence, but we should not ignore the impact of the excessive pursuit for a negative margin on the patient’s reproductive function protection and postoperative recovery. Long-term follow-up is necessary regardless of whether patients receive medical treatment or surgical treatment.</p

Table3_Characteristics and treatment strategies of aggressive angiomyxoma in women: A retrospective review of 87 cases.docx

ObjectiveAggressive angiomyxoma (AAM) is a rare kind of soft tissue tumor. The purpose of this study is to summarize the clinical manifestations and treatment strategy of AAM in women.MethodWe searched the case reports on AAM in EMBASE, Web of Science and PubMed, China biomedical database, Wanfang database, VIP database, and China National Knowledge Internet from the start of database construction to November 2022 without any language restrictions in place. Then, the obtained case data were extracted, summarized, and analyzed.ResultA total of 74 articles were retrieved involving 87 cases. The age ranges of onset were 2–67 years. The median age at onset was 34 years. The size of the tumor varied greatly among individuals, and about 65.5% of them were asymptomatic. MRI, ultrasound, and needle biopsy were used for diagnosis. Surgery was the primary mode of treatment, but it was prone to relapse. Gonadotropin-releasing hormone agonist (GnRH-a) might be used to reduce the tumor size before the operation and prevent recurrence after the operation. For patients who are unwilling to receive surgical treatment, GnRH-a alone could be attempted.ConclusionDoctors should consider the possibility of AAM in women with genital tumors. For surgery, it must be ensured that the negative surgical margin is recommended and achieved for preventing recurrence, but we should not ignore the impact of the excessive pursuit for a negative margin on the patient’s reproductive function protection and postoperative recovery. Long-term follow-up is necessary regardless of whether patients receive medical treatment or surgical treatment.</p

Table2_Characteristics and treatment strategies of aggressive angiomyxoma in women: A retrospective review of 87 cases.xlsx

ObjectiveAggressive angiomyxoma (AAM) is a rare kind of soft tissue tumor. The purpose of this study is to summarize the clinical manifestations and treatment strategy of AAM in women.MethodWe searched the case reports on AAM in EMBASE, Web of Science and PubMed, China biomedical database, Wanfang database, VIP database, and China National Knowledge Internet from the start of database construction to November 2022 without any language restrictions in place. Then, the obtained case data were extracted, summarized, and analyzed.ResultA total of 74 articles were retrieved involving 87 cases. The age ranges of onset were 2–67 years. The median age at onset was 34 years. The size of the tumor varied greatly among individuals, and about 65.5% of them were asymptomatic. MRI, ultrasound, and needle biopsy were used for diagnosis. Surgery was the primary mode of treatment, but it was prone to relapse. Gonadotropin-releasing hormone agonist (GnRH-a) might be used to reduce the tumor size before the operation and prevent recurrence after the operation. For patients who are unwilling to receive surgical treatment, GnRH-a alone could be attempted.ConclusionDoctors should consider the possibility of AAM in women with genital tumors. For surgery, it must be ensured that the negative surgical margin is recommended and achieved for preventing recurrence, but we should not ignore the impact of the excessive pursuit for a negative margin on the patient’s reproductive function protection and postoperative recovery. Long-term follow-up is necessary regardless of whether patients receive medical treatment or surgical treatment.</p

CREG1 heterozygous mice are susceptible to high fat diet-induced obesity and insulin resistance

<div><p><u>C</u>ellular <u>r</u>epressor of <u>E</u>1A-stimulated <u>g</u>enes 1 (CREG1) is a small glycoprotein whose physiological function is unknown. In cell culture studies, CREG1 promotes cellular differentiation and maturation. To elucidate its physiological functions, we deleted the <i>Creg1</i> gene in mice and found that loss of CREG1 leads to early embryonic death, suggesting that it is essential for early development. In the analysis of <i>Creg1</i> heterozygous mice, we unexpectedly observed that they developed obesity as they get older. In this study, we further studied this phenotype by feeding wild type (WT) and Creg1 heterozygote (<i>Creg1</i>^<i>+/-</i>) mice a high fat diet (HFD) for 16 weeks. Our data showed that <i>Creg1</i>^<i>+/-</i> mice exhibited a more prominent obesity phenotype with no change in food intake compared with WT controls when challenged with HFD. <i>Creg1</i> haploinsufficiency also exacerbated HFD-induced liver steatosis, dyslipidemia and insulin resistance. In addition, HFD markedly increased pro-inflammatory cytokines in plasma and epididymal adipose tissue in <i>Creg1</i>^<i>+/-</i> mice as compared with WT controls. The activation level of NF-κB, a major regulator of inflammatory response, in epididymal adipose tissue was also elevated in parallel with the cytokines in <i>Creg1</i>^<i>+/-</i> mice. These pro-inflammatory responses elicited by CREG1 reduction were confirmed in 3T3-L1-derived adipocytes with CREG1 depletion by siRNA transfection. Given that adipose tissue inflammation has been shown to play a key role in obesity-induced insulin resistance and metabolic syndrome, our results suggest that <i>Creg1</i> haploinsufficiency confers increased susceptibility of adipose tissue to inflammation, leading to aggravated obesity and insulin resistance when challenged with HFD. This study uncovered a novel function of CREG1 in metabolic disorders.</p></div

<i>Creg1</i>^<i>+/-</i> mice develop aggravated dyslipidemia, liver steatosis, glycaemia and insulin resistance on high fat diet (HFD).

<p>After 16 weeks of normal diet (ND) or HFD, (<b>A</b>): plasma lipid profile including total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL)-cholesterol and free fatty acid (FFA) of wild type (WT) and <i>Creg1</i>^<i>+/-</i> mice were measured by ELISA method (n = 8) and (<b>B</b>): liver steatosis was evaluated by H&E staining of paraffin-embedded sections (n = 8 to 10). (<b>C</b>): Fasting blood glucose, (<b>D</b>): fasting insulin concentration, and (<b>E</b>): homeostasis model assessment-insulin resistance [HOMA-IR = fasting glucose (mmol/l) × fasting insulin (mIU/l)/22.5] in WT and <i>Creg1</i>^<i>+/-</i> mice fed on ND or HFD for 16 weeks (n = 8). (<b>F</b>): Results of glucose tolerance test (GTT, upper panel) and quantification of area under curve (AUC) of GTT (lower panel) (n = 8). (<b>G</b>): Results of insulin tolerance test (ITT, upper panel) and quantification of inverse AUC of ITT (lower panel) (n = 8). Values are presented as means ± SEM. ns, no significance. *<i>P</i><0.05, **<i>P</i><0.01 and ***<i>P</i><0.001.</p

Reaction of [Ga2(tBu)4(neol-H)]2 with early transition metal chlorides and amides

Figure S4. Full-length cDNA sequence of TCD10 and mutation sites; The nucleotides with the blue (A), red (G) and green (CCATGGCCGGGTCGGGG) letters represent the deleted nucleotides in tcd10, T1-1 and T1-2 transgenic lines using CRISPR/Cas9 system technique; The sequences in the box represents the recognition sequences in CRIPPER/Cas9 experiments. (DOCX 21 kb