The authors used the following drugs at the following doses: mirogabalin (MIRO; 1, 10, 20, 40 mg/kg, MedChem Express, Monmouth Junction, NJ, USA), morphine hydrochloride (M; 5 mg/kg; Fagron, Krakow, Poland), buprenorphine (B; 5 mg/kg; Polfa S.A., Warsaw, Poland), oxycodone (O; 5 mg/kg; Norpharma A/S, Denmark), and ketamine (K; 15 mg/kg; Panpharma UK Ltd., Horsham, UK). The scheme of the experiments used in our current study were based on our previous published paper [48 (link)]. The dosages of morphine and buprenorphine were chosen based on our own studies [49 (link)], and the dosage of oxycodone was chosen based on our own unpublished data and literature [50 (link),51 (link)]. The dosage of ketamine was chosen based on the literature [52 (link)]. Before intraperitoneal administration (i.p.) to test mice, all the drugs were dissolved in water (aqua pro-injection). Vehicle (aqua pro-injection) was given to the control group according to the same protocol. During the study, no adverse effects of opioids or mirogabalin were noted.
Morphine hydrochloride
Manufactured by Fagron
Sourced in United States, Poland
Morphine hydrochloride is a pharmaceutical raw material used in the production of various medication formulations. It is a white crystalline powder that serves as the active pharmaceutical ingredient (API) in the manufacture of pain relief medications and other therapeutic products. The core function of morphine hydrochloride is to provide a reliable and standardized source of the opioid analgesic compound for use in medical applications.
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5 protocols using morphine hydrochloride
1
Opioid and Mirogabalin Dosage Comparison
2
Morphine and Enkephalin Protocol
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Morphine hydrochloride (20 µg/5 µL) (M; Fagron, Cracow, Poland) and UW1/enkephalin were used.
3
Intrathecal Administration of Bioactive Compounds
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The following compounds were used in our experiments: fisetin (F; Cayman, Ann Arbor, MI, USA), peimine (P; Sigma‒Aldrich, St Louis, USA), artemisinin (Ar; Sigma‒Aldrich, St. Louis, MI, USA), astaxanthin (A; Sigma‒Aldrich, St Louis, USA), bardoxolone methyl (BM, Sigma‒Aldrich, St. Louis, MI, USA), 740 Y-P (7; Med Chem Express, Monmouth Junction, NJ, USA), morphine hydrochloride (M; Fagron, Krakow, Poland), buprenorphine (B; Polfa S.A., Warsaw, Poland) and oxycodone hydrochloride (O; Molteni Farmaceutici, Scandicci, Italy). All of these substances were administered via intrathecal injection (i.t.), a standard procedure in our laboratory [109 (link),111 (link)]. It is performed by using a Hamilton syringe with a thin needle in accordance with instructions described previously [112 (link)]. The injections were performed in the lumbar segment of the spinal cord (between the L5 and L6 vertebrae) with a volume of 5 µL, and the tail reflex was an indicator of correct administration.
4
Synthesis and Characterization of Novel Pyrrolo[3,4-d]pyridazinone Derivatives
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The studied compounds, novel pyrrolo[3,4-d]pyridazinone derivatives, named 10b and 13b , were synthesized from commercially available reagents and solvents according to the previously described method [6 (link)]. The structures and purity of 10b and 13b were confirmed by different spectroscopic techniques: 1H NMR, 13C NMR, MS, FTIR, elemental analysis, and on the basis of their physicochemical properties.
Drugs and chemicals used in the experiment were as follows: Indomethacin (Sigma-Aldrich, Steinheim, Germany); morphine hydrochloride (Fagron, Kraków, Poland); carboxymethylcellulose (CMC) (PolAura, Olsztyn, Poland) dissolved to 0.5% solution in normal saline (Polpharma, Starogard Gdański, Poland); pentobarbital sodium + pentobarbital 133.3 mg/mL + 26.7 mg/mL, sol. (Morbital, Biowet, Puławy, Poland); medetomidine hydrochloride 1 mg/mL, sol. (Domitor, OrionPharma, Warszawa, Poland); formalin 37% sol. (Chempur, Piekary Śląskie, Poland) dissolved to 1.5% solution in normal saline (as above). Other chemicals used were included in the commercially available kits.
Drugs and chemicals used in the experiment were as follows: Indomethacin (Sigma-Aldrich, Steinheim, Germany); morphine hydrochloride (Fagron, Kraków, Poland); carboxymethylcellulose (CMC) (PolAura, Olsztyn, Poland) dissolved to 0.5% solution in normal saline (Polpharma, Starogard Gdański, Poland); pentobarbital sodium + pentobarbital 133.3 mg/mL + 26.7 mg/mL, sol. (Morbital, Biowet, Puławy, Poland); medetomidine hydrochloride 1 mg/mL, sol. (Domitor, OrionPharma, Warszawa, Poland); formalin 37% sol. (Chempur, Piekary Śląskie, Poland) dissolved to 1.5% solution in normal saline (as above). Other chemicals used were included in the commercially available kits.
5
Investigating Neuropathic Pain Treatments
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In the experiment, we used the following substances: SB328437 (SB), morphine hydrochloride (M; Fagron, Krakow, Poland), and buprenorphine (B; Polfa Warszawa S+A, Warsaw, Poland). SB328437 was dissolved in vehicle (V), whereas morphine and buprenorphine were dissolved in water for injection.
The first group of rats received a pretreatment with 10 μg/5 μl i.t. SB 16 and 1 h before CCI surgery and then once a day for the following 7 days. The control rats were administered vehicle (V) according to the same schedule. Behavioral tests (von Frey and cold plate tests) were performed on the 2nd and 7th days post-CCI 3 h after the vehicle or SB injection (Figure 4A
Moreover, on the 6th day after CCI surgery, other group of the chronically treated rats received a single dose of vehicle (V; water for injection) morphine (2.5 μg/5 μl, i.t.) or buprenorphine (1.0 μg/5 μl, i.t.) 30 min after V (70% DMSO) or CCR3 antagonist administration. These animals performed the same behavioral tests 30–35 min after V or opioid injection (it is 1 h after V or SB injections) (Figure 5A
The first group of rats received a pretreatment with 10 μg/5 μl i.t. SB 16 and 1 h before CCI surgery and then once a day for the following 7 days. The control rats were administered vehicle (V) according to the same schedule. Behavioral tests (von Frey and cold plate tests) were performed on the 2nd and 7th days post-CCI 3 h after the vehicle or SB injection (
Moreover, on the 6th day after CCI surgery, other group of the chronically treated rats received a single dose of vehicle (V; water for injection) morphine (2.5 μg/5 μl, i.t.) or buprenorphine (1.0 μg/5 μl, i.t.) 30 min after V (70% DMSO) or CCR3 antagonist administration. These animals performed the same behavioral tests 30–35 min after V or opioid injection (it is 1 h after V or SB injections) (
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