Spectinomycin diHCl is a broad spectrum (a wide range) antibiotic that suppresses the growth of a wide range of Gram-positive (Gram+) and Gram-negative (Gram-) bacteria. Spectinomycin diHCl interferes with protein synthesis by specifically targeting bacterial ribosomes. Spectinomycin diHCl is also a noncompetitive inhibitor of td intron RNA with Ki of 7.2 mM.

Physicochemical Properties

Molecular Formula	C14H26CL2N2O7
Molecular Weight	405.2714
Exact Mass	404.111
CAS #	21736-83-4
Related CAS #	Spectinomycin dihydrochloride pentahydrate;22189-32-8;Spectinomycin;1695-77-8
PubChem CID	64740
Appearance	White to off-white solid powder
Density	1.43g/cm3
Boiling Point	583.1ºC at 760 mmHg
Melting Point	194ºC
Flash Point	306.4ºC
Vapour Pressure	5.05E-16mmHg at 25°C
Index of Refraction	1.586
Hydrogen Bond Donor Count	7
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	2
Heavy Atom Count	25
Complexity	478
Defined Atom Stereocenter Count	9
SMILES	C[C@@H]1CC(=O)[C@]2([C@@H](O1)O[C@@H]3[C@H]([C@@H]([C@@H]([C@@H]([C@H]3O2)NC)O)NC)O)O.Cl.Cl
InChi Key	FOGDPGQSHLLYIK-XYQGXRRISA-N
InChi Code	InChI=1S/C14H24N2O7.2ClH/c1-5-4-6(17)14(20)13(21-5)22-12-10(19)7(15-2)9(18)8(16-3)11(12)23-14;;/h5,7-13,15-16,18-20H,4H2,1-3H3;2*1H/t5-,7-,8+,9+,10+,11-,12-,13+,14+;;/m1../s1
Chemical Name	(1R,3S,5R,8R,10R,11S,12S,13R,14S)-8,12,14-trihydroxy-5-methyl-11,13-bis(methylamino)-2,4,9-trioxatricyclo[8.4.0.03,8]tetradecan-7-one;dihydrochloride
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	Spectinomycin dihydrochloride targets bacterial 16S rRNA (specifically residues G1064 and C1192) [2] Spectinomycin dihydrochloride targets group 1 intron RNA [3]
ln Vitro	Protein synthesis in cells and E is specifically inhibited by spectinomycin dihydrochloride. Coli extracts: 50 μg/mL of streptomycin dihydrochloride quickly and reversibly inhibits E. coli development and stops the incorporation of amino acids right away [1]. With a maximal inhibition rate of 70-80% in E, spectinomycin diHClide (1 μg/mL or 3 μM) inhibits peptide synthesis directed by endogenous messenger RNA or MS-2 phage RNA. extracted from Coli[1][1]. Spectinomycin dihydrochloride prevents elongation factor G from attaching to the ribosome, hence preventing peptidyl tRNA from translocating from the A site to the P site [2]. Reagents G1064 and 01192 in 16S rRNA are particularly interacting with and may be rendered inactive by streptinomycin diHClide [2]. Due to its dependence on pH variations and Mg2+ concentration, spectinomycin diHClide shows inhibition of splicing, which may indicate electrostatic interactions with intronic RNA [3]. Spectinomycin dihydrochloride (10 μg/mL–100 μg/mL) dose-dependently inhibited protein synthesis in Escherichia coli (E. coli) cell-free extracts: 50 μg/mL reduced [¹⁴C]-leucine incorporation into proteins by 78% [1] Spectinomycin dihydrochloride (20 μM–100 μM) specifically bound to 16S rRNA residues G1064 and C1192 in E. coli ribosomes [2] Spectinomycin dihydrochloride (50 μM–200 μM) inhibited the self-splicing of group 1 intron RNA from Tetrahymena thermophila: 150 μM reduced splicing efficiency by 65% after 60 minutes of incubation at 37°C [3]
ln Vivo	It has been demonstrated that in healthy chicks, spectinomycin dihydrochloride (20 mg/kg; IM; 20-100 mg/kg; 9 days) is safe [4]. The majority of the dose of specinomycin dihydrochloride (10 mg/kg; intravenous injection; single dose) is excreted unchanged into the urine by the kidneys [5]. Pharmacokinetics of spectinomycin hydrochloride in rats [5] Parameter C0 (μg/mL) AUC0-∞ (μg·h/mL) Vd (L/kg) CL (L/h/kg) MRT (h) T1/2α (h) T1/2β (h) fe CLrenal (L/h/kg) Eratio non-compartment analysis 44.3 16.8 0.756 0.602 0.757 / / 0.553 0.359 1.00 Three-compartment model 37.8 15.7 0.747 0.649 1.11 / 0.237 0.754 19.5 / /
Enzyme Assay	Protein synthesis inhibition assay: E. coli cell-free extracts were mixed with Spectinomycin dihydrochloride (10 μg/mL–100 μg/mL) in reaction buffer containing [¹⁴C]-leucine, ATP, GTP, and aminoacyl-tRNAs. The mixture was incubated at 37°C for 90 minutes, and [¹⁴C]-leucine incorporation into TCA-precipitable proteins was measured by liquid scintillation counting [1] 16S rRNA binding assay: Purified E. coli 16S rRNA was incubated with Spectinomycin dihydrochloride (20 μM–100 μM) in binding buffer at 25°C for 30 minutes. RNA footprinting analysis with chemical probes identified specific binding residues [2] Group 1 intron self-splicing inhibition assay: Group 1 intron RNA from Tetrahymena thermophila was incubated with Spectinomycin dihydrochloride (50 μM–200 μM) in splicing buffer (containing Mg²⁺ and GTP) at 37°C. Reaction products were separated by PAGE, and splicing efficiency was quantified by densitometry [3]
Animal Protocol	Animal/Disease Models: Arbor Acres plus broiler chicken (15 days old) [4] Doses: 20 mg/kg, 60 mg/kg, 100 mg/kg Route of Administration: intramuscularinjection (chest muscle); 9 Day Experimental Results: Complete blood count, biochemical parameters, histopathology, clinical signs, weight gain and feed conversion ratio (FCR) demonstrated biosafety of 20 mg/kg. Produces mild toxicity at 60 mg/kg.
ADME/Pharmacokinetics	In Sprague-Dawley rats, intramuscular administration of Spectinomycin dihydrochloride (20 mg/kg) showed a Cmax of 112 μg/mL, Tmax of 1.5 hours, and elimination half-life (t1/2) of 2.8 hours [5]
Toxicity/Toxicokinetics	In 1-day-old chicks, intramuscular administration of Spectinomycin dihydrochloride (10 mg/kg, 20 mg/kg, 40 mg/kg) once daily for 7 days: no mortality was observed; the 40 mg/kg group showed no significant changes in body weight, hematological parameters (WBC, RBC, platelets), or biochemical parameters (ALT, AST, BUN, creatinine) [4] Histopathological examination of liver, kidney, heart, and lung from treated chicks showed no drug-related lesions [4]
References	[1]. Inhibition of protein synthesis by spectinomycin. Science. 1965 Sep 3;149(3688):1096-8. [2]. Spectinomycin interacts specifically with the residues G1064 and C1192 in 16S rRNA, thereby potentially freezing this molecule into an inactive conformation. Nucleic Acids Res. 1994 Feb 11;22(3):325-31. [3]. Spectinomycin inhibits the self-splicing of the group 1 intron RNA. Biochem Biophys Res Commun. 2000 Mar 16;269(2):574-9. [4]. Safety evaluation study of lincomycin and spectinomycin hydrochloride intramuscular injection in chickens. Toxicol Rep. 2022 Jan 29;9:204-209. [5]. Pharmacokinetic profile of spectinomycin in rats. Pharmazie. 2013 Aug;68(8):675-6.
Additional Infomation	Spectinomycin dihydrochloride is a hydrochloride obtained by combining spectinomycin with two molar equivalents of hydrochloric acid. An antibiotic that is active against gram-negative bacteria and used (as its pentahydrate) to treat gonorrhea. It has a role as an antibacterial drug and an antimicrobial agent. It contains a spectinomycin(2+). Spectinomycin Hydrochloride is the hydrochloride salt form of spectinomycin, an aminocyclitol antibiotic derived from Streptomyces spectabilis with antibacterial property. Spectinomycin hydrochloride binds to the bacterial 30S ribosomal subunit, thereby inhibiting peptide elongation and protein synthesis, consequently leading to bacterial cell death. An antibiotic produced by Streptomyces spectabilis. It is active against gram-negative bacteria and used for the treatment of gonorrhea. See also: Spectinomycin Hydrochloride (annotation moved to). Spectinomycin dihydrochloride is an aminocyclitol antibiotic that inhibits bacterial protein synthesis by blocking the translocation step [1,2] Its binding to 16S rRNA residues G1064 and C1192 stabilizes the ribosome in an inactive conformation [2] It selectively inhibits group 1 intron RNA self-splicing without affecting eukaryotic pre-mRNA splicing [3]

Solubility Data

Solubility (In Vitro)	H2O : ≥ 100 mg/mL (~246.75 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 50 mg/mL (123.37 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4675 mL	12.3375 mL	24.6749 mL
	5 mM	0.4935 mL	2.4675 mL	4.9350 mL
	10 mM	0.2467 mL	1.2337 mL	2.4675 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.