Discovery of a cell-active chikungunya virus nsP2 protease inhibitor using a covalent fragment-based screening approach.

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that has been responsible for numerous large-scale outbreaks in the last twenty years. Currently, there are no FDA-approved therapeutics for any alphavirus infection. CHIKV non-structural protein 2 (nsP2), which contains a cysteine protease domain, is essential for viral replication, making it an attractive target for a drug discovery campaign. Here, we optimized a CHIKV nsP2 protease (nsP2pro) biochemical assay for the screening of a 6,120-compound cysteine-directed covalent fragment library. Using a 50% inhibition threshold, we identified 153 hits (2.5% hit rate). In dose-response follow up, RA-0002034, a covalent fragment that contains a vinyl sulfone warhead, inhibited CHIKV nsP2pro with an IC 50 of 58 ± 17 nM, and further analysis with time-dependent inhibition studies yielded a k inact /K I of 6.4 x 10 3 M -1 s -1 . LC-MS/MS analysis determined that RA-0002034 covalently modified the catalytic cysteine in a site-specific manner. Additionally, RA-0002034 showed no significant off-target reactivity against a panel of cysteine proteases. In addition to the potent biochemical inhibition of CHIKV nsP2pro activity and exceptional selectivity, RA-0002034 was tested in cellular models of alphavirus infection and effectively inhibited viral replication of both CHIKV and related alphaviruses. This study highlights the discovery and characterization of the chemical probe RA-0002034 as a promising hit compound from covalent fragment-based screening for development toward a CHIKV or pan-alphavirus therapeutic. Significance Statement: Chikungunya virus is one of the most prominent and widespread alphaviruses and has caused explosive outbreaks of arthritic disease. Currently, there are no FDA-approved drugs to treat disease caused by chikungunya virus or any other alphavirus-caused infection. Here, we report the discovery of a covalent small molecule inhibitor of chikungunya virus nsP2 protease activity and viral replication of four diverse alphaviruses. This finding highlights the utility of covalent fragment screening for inhibitor discovery and represents a starting point towards the development of alphavirus therapeutics targeting nsP2 protease.

Young adults with spina bifida may have higher occurrence of prehypertension and hypertension.

OBJECTIVE: The aims of this study were to determine the occurrence of prehypertension and high blood pressure in adults with spina bifida (SB) and to examine relationships among blood pressure, cardiovascular disease risk factors, and SB-specific factors. DESIGN: This is a cross-sectional, retrospective analysis of adults with SB. SB-specific factors and cardiovascular disease risk factors were compared among subjects with high blood pressure, subjects with blood pressure in the prehypertensive range, and normotensive subjects using the χ, Kruskal-Wallis, or Fisher's exact tests. RESULTS: Of 225 subjects, the occurrence of prehypertension and high blood pressure was 27% (n = 22) and 27% (n = 22) for ages 18-29 yrs, 35% (n = 26) and 41% (n = 30) for ages 30-39 yrs, 18% (n = 8) and 66% (n = 29) for ages 40-49 yrs, and 21% (n = 5) and 67% (n = 16) for 50 yrs or older, respectively. Of the subjects with high blood pressure, 56% were men, 14% had obstructive sleep apnea, 14% had diabetes, 19% had renal dysfunction, 38% used tobacco, 16% had hydronephrosis, and 71% had a shunt. The groups differed significantly with respect to diabetes (P = 0.004), bladder procedures (P = 0.001), and renal dysfunction (P < 0.001), with higher proportions of subjects with high blood pressure having these comorbidities. CONCLUSIONS: Fewer than half of the subjects were considered normotensive. A greater proportion of young adults with SB appear to have high blood pressure compared with the general United States population. Because elevated blood pressure is an independent, modifiable risk factor of cardiovascular disease, these findings support early screening and intervention for elevated blood pressure in individuals with SB.

Optimizing health care for adults with spina bifida.

Survival into adulthood for individuals with spina bifida has significantly improved over the last 40 years with the majority of patients now living as adults. Despite this growing population of adult patients who have increased medical needs compared to the general population, including spina bifida (SB)-specific care, age-related secondary disabilities, and general adult medical needs, there is little published information about the natural history of SB in adulthood. There are few published studies of medical conditions, interventions, or long-term complications in this population. This article will provide a review of the medical issues of adults with SB, highlighting areas that are different than pediatric care, and areas of needed research.

Medical care of adults with spina bifida.

Survival into adulthood for individuals with spina bifida has significantly improved over the last 40 years. Health services research suggests the majority of patients with spina bifida are now over 18 years old. Adults with spina bifida have ongoing increased medical needs compared to the general population, including spina bifida-specific care, age-related secondary disabilities, and general adult medical needs. Unlike pediatric-aged patients, adults may not have access to multi-disciplinary spina bifida clinics and must often coordinate their own care with adult providers less familiar with spina bifida and the associated monitoring and treatment issues. This article will provide an overview of the medical issues of adults with spina bifida, highlighting areas that are different than pediatric care.