Unveiling the Chemistry and Applications of Flakka USAID Business Growth Activity

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 alpha-pyrrolidinopentiophenone function 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.

Effects of PV8, 4-F-PV8, and 4-MeO-PV8 on the Survival of SH-SY5Y, Hep G2, RPMI 2650, and H9c2(2- Cells

Most previous work with stimulant-induced neurotoxicity has focused on substrate-based releasers rather than reuptake inhibitors. Moreover, hyperthermic responses to reuptake inhibitors have been less forthcoming than with substrate-based releasers, and we did not observe a hyperthermic response to cocaine in the present study. In the current study, we assessed the persistent behavioral and neurochemical effects of exposure to the second-generation synthetic cathinone α-pyrrolidinopropiophenone (α-PPP). Both 4MMC and α-PVP produce robust functional changes in the DA system, albeit by different mechanisms of action, releasing DA, or blocking DA transport, respectively (Baumann et al., 2012; Cameron et al., 2013; Glennon and Young, 2016; Marusich et al., 2014; Simmler et al., 2013). Overall, there were few neurochemical differences following self-administration of a DA uptake inhibitor versus a DA releaser, and the magnitude of the differences between effects of these synthetic cathinones was small (Fig. 3 and 4). This suggests that synthetic cathinones may lead to similar neuroplasticity, regardless of the mechanism of action.

Unveiling the Chemistry and Applications of Flakka

Drug concentrations used in this in vitro study, reaching 300 μM, exceed those normally found in the blood obtained from intoxicated patients and during autopsies (Kudo et al. 2015; Marinetti and Antonides 2013). However, as discussed in our previous work (Wojcieszak et al. 2016), organs such as the liver, brain, and upper airway epithelium can be exposed to significantly higher local drug concentrations than those measured in blood. Moreover, it is noteworthy that immortalized cancer cell lines, which are a convenient model for in vitro studies, can be more resistant to cytotoxicity, and therefore, cell damage can be observed in concentrations higher than in normal cells in vivo (den Hollander et al. 2014; Wojcieszak et al. 2016).

• The report demonstrates, for the first time, that changes of fluidity of the interior part of plasma membrane contribute to the cytotoxicity of pyrovalerone derivatives, in addition to the previously reported mechanisms.
• Thus, it is unlikely that the timing of the analyses caused differences in neurotransmitter levels.
• These decrements were also selective, as there were no changes in dopamine levels or spatial memory (Lopez-Arnau et al. 2014).
• Pyrovalerone derivatives (α-pyrrolidinophenones) form a branch of synthetic cathinones, a second most prominent group of novel psychoactive substances.
• Behavior was assessed 4–5 days after the dosing regimen, and neurochemistry was assessed the following day.
• Some Flakka abusers have reported dissociative feelings similar to those felt when abusing benzodiazepines or hallucinogens such as ketamine.

Spontaneous alternation in the Y-maze has been proposed to measure hippocampus-dependent spatial working memory (Walker and Gold 1994). Performance in the Y-maze was assessed by both automated quantitation (Maze Engineers; Cambridge, MA) and manual observation. Spontaneous alternations were calculated as the percentage of the total arm entries minus two composed of triads containing entries into all arms. For α-PVP, 4-MEC, and METH, higher doses (5, 100, and 3mg/kg, respectively) produced elevations in ICSS threshold values, with only METH producing significant elevations (mean±95% CI; α-PVP, 19.83±38.64; 4-MEC, 28.00±31.72; METH, 84.39±69.48%).

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.

What happened to the “Man on Flakka?”

Statistical analyses were conducted using NCSS (Number Cruncher Statistical Systems, Kaysville, Utah, USA). In most instances, data from naïve and LgA groups (present study) were statistically analyzed separately from ShA groups (Marusich et al., 2019a; Marusich et al., 2019b) because the neurotransmitter assays were conducted at different times for these studies. Additionally, one male 4MMC ShA and saline ShA rat were exposed to the incorrect infusion volume.

Fig. 5.

Incubation of cells with 4-F-PVP for 72 h resulted in a marked, concentration-dependent decline in the survival of all cell lines, with the maximal effect at the similar level in SH-SY5Y (reduction by 60%), Hep G2 (reduction by 54%), RPMI 2650 (reduction by 59%), and H9c2(2-1) (reduction by 46%) (Fig. 2b). Α-PPP exposure results in persistent changes in exploratory behavior, spatial working memory, and monoamine neurochemistry. This research highlights potential dangers of α-PPP, including potential neurotoxicity, and suggests that the mechanisms underlying the persistent untoward effects of the cathinones may be distinct from those of the amphetamines.

LgA self-administration of α-PVP increased 5-HIAA levels in all brain regions, compared to control. In contrast, both LgA and ShA 4MMC self-administration decreased 5-HT and 5-HIAA levels in most brain regions. LgA exposure to both synthetic cathinones increased DOPAC levels in hypothalamus and striatum, and increased HVA levels in striatum compared to control.

• Specifically, synthetic cathinone products have been shown to often contain more than one cathinone, as well as other adulterants, including illicit amphetamines, piperazines, cutting/binding agents, caffeine, and topical anesthetics (Brandt et al., 2010; German et al., 2014).
• Incubation of cells with 4-F-PVP for 72 h resulted in a marked, concentration-dependent decline in the survival of all cell lines, with the maximal effect at the similar level in SH-SY5Y (reduction by 60%), Hep G2 (reduction by 54%), RPMI 2650 (reduction by 59%), and H9c2(2-1) (reduction by 46%) (Fig. 2b).
• Statistical analyses were conducted using NCSS (Number Cruncher Statistical Systems, Kaysville, Utah, USA).
• Thus, while α-PVP and 4MMC are known to elevate DA (Baumann et al., 2012; Baumann et al., 2017; Kehr et al., 2011; Marusich et al., 2014), this affect is fleeting, and was not present one day after final drug exposure when brain samples were collected.
• One limitation is that the neurotransmitter data for LgA and naïve were analyzed at a different time than those for ShA.
• These findings are consistent with a past study showing that ShA 4MMC self-administration did not alter DA, DOPAC, or HVA in striatum (Motbey et al., 2013).
• Results obtained using LDH assay further confirm the impact of the side-chain length on the cytotoxicity of pyrrolidinophenones.

Long incubation times were applied in order to show whether the cytotoxicity of studied compounds increase with time, which is relevant since the common abuse pattern of synthetic cathinones includes long sessions during which multiple doses are administered (Zawilska and Wojcieszak 2013). The effects of α-PPP on serotonin levels are somewhat surprising, as it has reported selectivity for the dopamine and norepinephrine transporters over the serotonin transporter (Eshleman et al. 2017). However, we used a relatively high dose regimen to ensure near maximal levels of toxicity, and α-PPP likely loses some selectivity at such doses.

This condition can lead to psychotic episodes that include violence towards others and self-injury. In the mid-2010s, police believe a man who murdered a couple and attempted to eat the male victim’s face was high on Flakka. Effective treatment typically includes a combination of medical detoxification, behavioral therapies, and ongoing support to manage cravings and prevent relapse. Law enforcement and community activists were instrumental in limiting the damage done by the drug’s dangerous effects. Most reports of flakka after 2016 turned out to be other similar chemicals in the bath salt family. Flakka is a dangerous drug that has many bad side effects, mostly including changes in behavior or mood.