Unveiling the Chemistry and Applications of Flakka USAID Business Growth Activity

Importantly, our results suggest that the risk of intoxication with pyrovalerones, resulting from their cytotoxic properties, could be positively related to the length of their aliphatic side-chain. On January 22, 2013, the US DEA published a request for information specifically regarding 8 additional synthetic cathinones, 2 of the most prominent being 4-methyl-N-ethcathinone (4-MEC) and α‐pyrrolidinopentiophenone (α-PVP) (DEA, 2013). Their similarity in chemical structure suggests that α-PVP and 4-MEC likely emerged as replacements for MDPV and mephedrone, respectively (Figure 1). Rats were surgically implanted with chronic indwelling jugular catheters under general anesthesia as previously described (Marusich et al., 2019a; Marusich et al., 2019b). Catheters were flushed daily with saline prior to the session, and with 0.2 ml of a solution containing 0.96% gentamicin, 2.88% heparin, and 96.2% saline after the session to maintain patency.

Flakka (Alpha-PVP) Overview, Effects, Abuse, Addiction, & Treatment

This study sought to examine the behavioral and neurochemical effects of extended access to α-pyrrolidinopentiophenone (α-PVP) and 4-methylmethcathinone (4MMC). Male and female Sprague-Dawley rats were trained to self-administer α-PVP (0.1 mg/kg/infusion) or 4MMC (0.5 mg/kg/infusion) through autoshaping, and then self-administered for 21 days during 1 h (short access; ShA) or 6 h (long access; LgA) sessions. Amygdala, hippocampus, hypothalamus, prefrontal cortex (PFC), striatum, and thalamus were extracted, and tissue was analyzed with electrochemical detection and liquid chromatography mass spectrometry. Rats acquired self-administration of α-PVP and 4MMC, and LgA rats showed more escalation of self-administration than ShA rats.

Sex differences for α-PVP during autoshaping (Fig. S2) did not lead to sex differences in α-PVP self-administration (Fig. 2). LgA α-PVP groups showed sex differences in self-administration on one day (Fig. 1), but this effect did not persist into other days of the study. Thus, the similarity in α-PVP intake across sexes was akin to that found in previous studies (Aarde et al., 2015; Javadi-Paydar et al., 2018). The similarity in 4MMC self-administration for both sexes is also consistent with past studies, which showed no sex difference in acquisition of 4MMC self-administration (Creehan et al., 2015; Vandewater et al., 2015). Sex differences in ShA saline self-administration were noted (Fig. 2) but were only statistically different for two days. The dosing consisted of a regimen of 4 unit doses of 80 mg/kg of α-PPP or saline administered over an 8 hour period with each dose separated by 2 hours.

Taken together, we predict that α-PVP possesses a potential for compulsive abuse (ie, addiction) that is roughly similar to that of METH and MDPV but much greater than that of 4-MEC, methylone, and MDMA. Accordingly, we also predict that 4-MEC will have a relatively lower potential for compulsive use than that of MDPV and METH, would be most similar to methylone and MDMA (ie, episodic use), and may exert primarily entactogenic effects. Rats were euthanized by alpha-pyrrolidinopentiophenone function rapid decapitation approximately 24 h after their last self-administration session in order to avoid direct drug effects (Marusich et al., 2019a; Marusich et al., 2019b; Wee et al., 2007b). The brain was then cut down the mid line and each cortical half was opened, and the hippocampus was removed (Spijker, 2011). Next, each half was cut into three coronal slices using midline anatomical markers moving rostral to caudal.

• Significant reductions were also seen in H9c2(2-1) cells between 10 and 300 μM, and in SH-SY5Y, Hep G2, and RPMI 2650 cells between 25 and 300 μM after 72-h incubation (Fig. 6b).
• Its molecular pharmacology was recently characterized, demonstrating that α-PPP has high affinity for the dopamine and norepinephrine transporters (approximately 1–2 μM), where it functions as a reuptake inhibitor.
• Finally, it is also important to mention that both first- and second-generation synthetic cathinones are often sold as mixtures.
• Behavior was studied using the elevated plus maze, Y-maze, and novel object recognition tests.
• Muscle tissue begins to break down, releasing proteins and other cellular products into the bloodstream, in a process called rhabdomyolysis.
• The effect was always significant at 200 and 300 μM for all drugs and cell lines; in addition, the membrane integrity of RPMI 2650 cells was also significantly affected by all drugs at 100 μM.
• It is an ongoing challenge, as each time one type of bath salt is made illegal, the drug labs change the chemical structure slightly and a new drug that is technically not illegal is created.

Similarly, after 24-h incubation, 4-MeO-PV8 decreased the viability of all tested cell lines (25–300 μM for SH-SY5Y and RPMI 2650 cells, 200 and 300 μM for Hep G2 and H9c2(2-1) cells), with the greatest effect being 59% reduction of the survival for SH-SY5Y, 68% for Hep G2, 87% for RPMI 2650, and 33% for H9c2(2-1). Extending incubation time to 72 h increased the cytotoxicity at 300 μM, leading to the decrease of the viability by 91% for SH-SY5Y, 97% for Hep G2, 98% for RPMI 2650, and 63% for H9c2(2-1). Moreover, a broader concentration range was found to elicit a significant drop in the viability for the Hep G2 (25–300 μM) and H9c2(2-1) (10–300 μM) cell lines, compared to 24-h exposure (Fg. 4c). Cell viability and mitochondrial function were measured by assessment of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reduction by mitochondrial dehydrogenases after 24- and 72-h exposure to the drugs. A solution of MTT (0.5 mg/ml) was added to the cells, and the culture was incubated for a further 3-h at 37 °C.

Mental Health: Substance Abuse in Older Adults

Following governmental bans of methylenedioxypyrovalerone, mephedrone, and methylone, a second generation of synthetic cathinones with unknown abuse liability has emerged as replacements. One limitation is that the neurotransmitter data for LgA and naïve were analyzed at a different time than those for ShA. This difference in analyses is a potential reason for the neurochemical differences between LgA and ShA groups. GLU levels were similar to the original analyses (Supplementary Material Table S2), and the differences between LgA and ShA were still evident in the reanalysis.

Treatment for Flakka abuse

• One of the most problematic classes of novel psychoactive substances to emerge are synthetic cathinones, comprising approximately 18% of all unregulated substances in international markets (United Nations, 2013).
• The brain was then cut down the mid line and each cortical half was opened, and the hippocampus was removed (Spijker, 2011).
• A prototypical second-generation pyrrolidine synthetic cathinone is the compound alpha-pyrrolidinopropiophenone (α-PPP).
• For the locomotor activity and core temperature experiments, an additional six mice were prepared with telemetry probes (see surgery section).
• Self-administration of either synthetic cathinone also increased NE levels in several brain regions, with the biggest increases shown in hypothalamus.
• The plate was mixed at 1000 RPM for 4 min on an Eppendorf MixMate orbital shaker (Enfield, CT).
• The onset of a Flakka overdose is often rapid and can cause heart problems, aggressive behavior, and psychosis.

Brain tissue samples were weighed and placed in cryovials containing stainless steel grinding balls. Supernatant was then transferred to a 96 well plate for analysis by electrochemical detection (ECD). A Thermo Scientific CoulArray Multi-Channel ECD Array system (model 5600A; Thermo Scientific, Waltham, MA, USA) was used to analyze neurotransmitter concentrations. The array detector contained 16 coulometric electrochemical cells that provided quantitation of multiple neurotransmitters and metabolites simultaneously.

Studies

Second, an unknown peak interfered with the integration of the DOPAC or NE peak in select samples of PFC and hypothalamus. Additionally, in some samples of thalamus, DOPAC levels were below the level of quantitation. Instead of omitting these values from graphs and analyses, we substituted a value of 0.005 ng/mg, which is halfway between zero and the level of quantitation of 0.01 ng/mg. For self-administration data, autoshaping data were analyzed with separate mixed factors ANOVAs (day x lever x sex) to compare active and inactive lever presses, with day and lever as within-subjects factors, and sex as a between-subjects factor. The additional 21 days of self-administration were also analyzed with separate mixed factors ANOVAs (day x lever x sex), which compared responses on the active and inactive levers. If the assumption of circularity was violated, the Geisser-Greenhouse Adjustment was employed.

Drug users take flakka to get a feeling of euphoria, a heightened sense of awareness, stimulation, and energy. Word on the street is that flakka (also called gravel or flocka) is a combination of heroin and crack cocaine, or heroin and methamphetamines, but in reality, flakka is just a newer-generation version of a type of synthetic drug called bath salts (MDPV). Law enforcement agencies and health care providers have expressed significant concerns about the widespread use of flakka, particularly among younger demographics and in clubbing scenes where synthetic drugs are more prevalent. If you or someone you know needs assistance, consider reaching out to a professional or a helpline.

Materials and Methods

Although spontaneous alternation behavior has been closely linked to brain cholinergic systems (Lalonde 2002), previous studies have documented a role for catecholamines, as dopamine depletions in the striatum or septum result in impaired spontaneous alternations (Taghzouti et al. 1985). Likewise, norepinephrine levels in the hippocampus are elevated during spontaneous alternations (Men et al. 1999) and depletion of norepinephrine from forebrain projections results in impaired spontaneous alternation (Pisa and Fibiger 1983). Relating specific neurochemical depletions to specific behavior impairments presents rich research opportunities. In this study, we document persistent effects of the second-generation pyrrolidine synthetic cathinone α-PPP on behavior and neurochemistry. We document that, consistent with previous studies of amphetamine derivatives (Murnane et al. 2012), α-PPP acutely decreases body weight over the course of a standard 6-hour dosing regimen.

PVP and its fluoro- and methoxy-substituted derivatives produced moderate and concentration- and time-dependent decline in the viability of SH-SY5Y, Hep G2, RPMI 2650, and H9c2(2-1) cells measured as mitochondrial activity. Cellular membrane integrity was assessed by measuring the activity of lactate dehydrogenase released from damaged cells into the culture medium using LDH Cytotoxicity Assay (ScienCell Research Laboratories, Carlsbad, CA, USA) following 48-h exposure to the drugs, according to the manufacturer’s instruction. Each experiment included a positive control of 1% (v/v) Triton-X100, as recommended by the manufacturer. Results are expressed as a percentage value of the positive control group, considered as 100% cytotoxicity. All graphical data presentations were created using GraphPad Prism 7.0 (GraphPad Software Inc.; La Jolla, CA). The time courses of the telemetry data (locomotor activity and core temperature) comparing saline versus α-PPP were integrated by standard area under the curve analysis in GraphPad, and then analyzed by repeated measures one-way analysis of variance, with post-hoc comparisons by Tukey’s test.