Mucuna pruriens Detoxification through Ensiling

Mucuna pruriens is grown for food and feed despite its L-Dopa concentration (2-6% by wt), which is toxic to monogastrics. The aim of this study was to determine if the L-Dopa concentration of Mucuna could be reduced by ensiling. The objective of Experiment 1 was to examine how long it takes to decrease the pH of ensiled Mucuna to 4.5. Crushed Mucuna beans (6 mm) were ensiled in the dark at room-temperature (18 to 25°C) for 0, 3, 7, 14, 21, and 28 days in vacuum-sealed bags. A pH of 4.5 and an L-Dopa concentration of 1.3% (54%) reduction) were recorded after 28 days. The objective of Experiment 2 was to study the effect of particle size of ensiled Mucuna on L-Dopa concentration and fermentation and nutritional characteristics. Ensiling Mucuna that had been ground to particle sizes of 2, 4 and 6 mm for 28 days decreased the L-Dopa concentration from 2.8%) in the unesiled bean to 1.2, 1.6, and 1.1%, respectively. Ensiling also reduced the water-soluble carbohydrate (WSC) concentration and pH and increased the ammonia-N (NH3N) concentration. Neither ensiling nor particle size affected concentrations of ether extract (EE; 5%), CP (23-25%), starch (38-40%), and neutral detergent fiber (NDF; 17- 20%). Dry matter losses (< 1%) and mold or yeast counts were unaffected by particle size. Aerobic stability was maintained beyond 657 hours in all treatments and the lactate:acetate ratio of all samples exceeded 3.0. In conclusion, ensiling Mucuna bean for 28 days reduced the L-Dopa concentration by 43 to 61% while preserving most nutrients. Particle size had minimal effects on nutritional composition or fermentation indices

Effect of Sonication and Two Solvent Extraction Methods on the L-dopa Concentration and Nutritional Value of Mucuna pruriens

Mucuna pruriens beans are high in crude protein (CP; 25-30%) and starch (39-41%), but also contain toxic L-Dopa. Methods to reduce L-Dopa to a safe threshold (< 0.4%) are often costly and little is known about their impact on the nutritional value of the bean. The objective of this study was to examine effects of three extraction methods on L-Dopa concentration and nutritional composition of finely (1 mm) or coarsely (6 mm) ground beans. Methods evaluated included extraction in solutions of acetic acid (ACD, pH 3) or sodium hydroxide (ALK, pH 11) for 8 hours or sonication (SON) for 5 minutes. All extraction methods decreased the L-Dopa concentration of fine Mucuna particles from 2.8 to < 0.2%, decreased their CP and water-soluble carbohydrate (WSC) concentrations by 24-31% and 78-81%, respectively and increased their NDF and starch concentrations by at least 62 and 14%, respectively. Sonication reduced the ether extract (EE) concentration of fine particles from 5.5% to 4.2% but ACD and ALK did not. Sonication and ACD did not reduce the L-Dopa concentration of coarsely ground beans but ALK reduced it from 2.8 to 2%. Sonication also reduced CP, WSC, and ether EE concentrations of coarse particles by 6, 17, and 27%, respectively and ALK increased their starch concentration by 17%. Therefore, the extraction methods reduced L-Dopa of fine Mucuna particles to safe levels but increased their NDF and starch concentrations at the expense of WSC and CP concentrations. Extraction methods were less effective at reducing the L-Dopa in coarse particles and had fewer, less consistent effects on their nutritional composition

Physiological and Performance Effects on Rats Fed Detoxified Mucuna pruriens

L-Dopa (3, 4 dihydroxy-L-phenylalanine), a toxic compound in Mucuna pruriens, causes reduced feed intake, anorexia, diarrhea, vomiting, skin lesions and potential mortality when consumed by humans and monogastric livestock. Processing the bean can reduce L-Dopa in Mucuna beans to safe levels (< 0.4%), but few studies have examined the effects of feeding detoxified Mucuna to monogastrics. The objective of this study was to examine the effect of feeding detoxified Mucuna bean on the performance, behavior, and health of rats. Sixty Sprague-Dawley rats were randomly assigned to five treatments (n=12). Dietary treatments consisted of a commercial rat chow (CON) or diets in which 10% of a customized rat chow was replaced with either undetoxified Mucuna (MUC), or Mucuna detoxified by acetic acid extraction (pH 3), sodium hydroxide extraction (pH 11), or ensiling for 28 days (SIL). During the 14-day trial, behavior, physiological development, and signs of clinical pathology were evaluated. Necropsy revealed that MUC caused splenomegaly and monocytosis, and reduced phosphorus concentrations relative to CON. These effects were not observed in rats fed detoxified diets, which alkaline phosphatase concentrations 11-17% greater than those of MUC, but similar to those of CON. No abnormalities in behavior, performance, or physiology were observed in any of the rats on the detoxified diets. Compared to those fed CON, rats fed Mucunabased diets had similar feed intake, weight gain, and behavioral results in the open field. It can be concluded that at the 10% level of dietary inclusion, there were fewer measurable adverse effects due to feeding the detoxified Mucuna bean compared to untreated Mucuna bean

Abstract PD11-09: PD11-09 Datopotamab deruxtecan (Dato-DXd) + durvalumab (D) as first-line (1L) treatment for unresectable locally advanced/metastatic triple-negative breast cancer (a/mTNBC): updated results from BEGONIA, a phase 1b/2 study

Abstract Background: Patients with a/mTNBC have limited treatment options and a poor prognosis (objective response rate [ORR] of 37%, median duration of response 6.5 months, median overall survival 15.5 months for 1L chemotherapy [Rugo, et al. Ann Oncol. 2021 LBA16]). Combining checkpoint inhibitors with 1L chemotherapy modestly improves outcomes but only in PD-L1–positive a/mTNBC, emphasizing a critical unmet need for patients with PD-L1–negative disease and for further improving outcomes in PD-L1–positive disease. BEGONIA (NCT03742102) is an ongoing 2-part, open-label platform study, evaluating safety and efficacy of D, an anti–PD-L1 antibody, combined with other novel therapies in 1L a/mTNBC, including Dato-DXd, an antibody-drug conjugate consisting of a humanized anti-TROP2 antibody covalently linked to a highly potent topoisomerase I inhibitor payload via a stable, tumor-selective, tetrapeptide-based cleavable linker. Early data from BEGONIA of D in combination with Dato-DXd showed promising responses. Here, we report updated results of Dato-DXd + D. Methods: Patients with unresectable a/mTNBC eligible for 1L treatment were enrolled, regardless of PD-L1 or TROP2 status, and received intravenous Dato-DXd 6 mg/kg + D 1120 mg every 3 weeks until progression or unacceptable toxicity. PD-L1, assessed using the VENTANA PD-L1 (SP263) Assay, was defined as high if ≥ 5% of the tumor area was populated by PD-L1–expressing tumor or immune cells. Primary endpoints were safety and tolerability. Secondary endpoints included investigator-assessed ORR (RECIST v1.1) and duration of response. Patients included in the efficacy analysis had ≥ 2 on-treatment disease assessments, progressed, died, or withdrew from the study. Results: As of April 8, 2022, 47 patients received Dato-DXd + D (39 ongoing) and 33 of those were included in the efficacy analysis. Median (range) follow-up was 7.5 (0–11) months. Patient age was a median of 51 years, 57% received prior treatment for early stage TNBC, and 60% had visceral metastases at baseline. Confirmed ORR was 26/33 (79%; 95% CI, 61–91); 2/33 patients (6%) had a complete response and 24/33 (73%) had a partial response. Confirmed response was irrespective of PD-L1 expression (PD-L1 high ORR, 4/5 [80%]; PD-L1 low, 16/21 [76%]; PD-L1 missing, 6/7 [86%] patients). Median duration of response was not reached; 100% of patients with a complete or partial response remained in response at 6 month follow-up, and 96% had an ongoing response at data cutoff. Adverse events (AEs) were manageable and consistent with the known safety profiles of each agent, with treatment-related AEs occurring in 41 patients (87%), any Grade 3/4 AEs in 17 patients (36%), and any serious AEs in 7 patients (15%). The most common all-Grade AEs were gastrointestinal (nausea in 26 patients [55%] and stomatitis in 24 patients [51%]). A low rate of diarrhea was reported (6 patients [13%], all Grade 1 or 2); 4 patients had anemia and 1 had neutropenia. There were no cases of interstitial lung disease/pneumonitis or thrombocytopenia. Nine patients (19%) and 11 patients (23%) underwent Dato-DXd dose reduction and delay, respectively; 14 (30%) had D dose delay. Treatment was discontinued due to an AE for 3 patients (6%). There were no deaths due to treatment-related AEs. Conclusions: In this updated analysis with additional patients and longer follow-up, the combination of Dato-DXd + D in 1L a/mTNBC demonstrated a manageable safety profile and compelling high response rates with promising durability. Although subgroups were small, responses occurred irrespective of PD-L1 expression. Further investigation of this treatment combination is warranted. Analysis of translational data is ongoing. Funding: AstraZeneca/Daiichi Sankyo Citation Format: Peter Schmid, Piotr Wysocki, Cynthia Ma, Yeon H. Park, Ricardo Fernandes, Simon Lord, Richard D. Baird, Catherine Prady, Kyung Hae Jung, Jamil Asselah, Robert Huisden, Ross Stewart, Petra Vuković, Ana T. Nunes, Zbigniew Nowecki. PD11-09 Datopotamab deruxtecan (Dato-DXd) + durvalumab (D) as first-line (1L) treatment for unresectable locally advanced/metastatic triple-negative breast cancer (a/mTNBC): updated results from BEGONIA, a phase 1b/2 study [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD11-09.</jats:p