Broad spectrum aminoglycoside antibiotic

In Caco-2 cells, plasmomycin sulfate (500 μg/ml) decreased intracellular parasitic forms by 97.2% and intracellular C by 99.5%. In HCT-8 cells as opposed to control, parvum forms. %[2].

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The structure of a cytosine–cytosine (CC) mismatch-containing RNA molecule derived from a hairpin structure in the thymidylate synthase mRNA that binds the aminoglycoside Paromomycin with high affinity was determined using nuclear magnetic resonance (NMR) spectroscopy. The cytosines in the mismatch form a noncanonical base pair where both cytosines are uncharged and stack within the stem of the RNA structure. Binding to paromomycin was analyzed using isothermal titration calorimetry (ITC) to demonstrate the necessity of the CC mismatch and to determine the affinity dissociation constant of this RNA to Paromomycin to be 0.5 ± 0.3 μM. The CC mismatch, and the neighboring GC base pairs experienced the highest degree of chemical shift changes in their H6 and H5 resonances indicating that paromomycin binds in the major groove at the CC mismatch site. In comparing the structure of CC mismatch RNA with a fully Watson–Crick GC base paired stem, the CC mismatch is shown to confer a widening of the major groove. This widening, combined with the dynamic nature of the CC mismatch, enables binding of paromomycin to this RNA molecule [1].

Oocysts per gram of feces and intestinal tissue are decreased with pomomycin sulfate (oral gavage; 50 mg/kg-200 mg/kg; once day; for five days, two weeks after infection). Just 20% of sections in the mice's intestines at the 50 mg/kg dose and 10% at the 200 mg/kg dose, respectively, displayed mild localized inflammation due to Cryptosporidium parvum infection. Inflammation in the foci.

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Cryptosporidium parvum is a protozoan parasite that infects the gastrointestinal epithelial cells causing several parasitological and pathological changes. It is incriminated in the development of colorectal cancer in immunosuppressed individuals. This study aimed to evaluate the effectiveness of low and high doses of Pparomomycin sulfate in the treatment of cryptosporidiosis in mice. Five groups of mice were included: group I, infected control; group II, infected and immunosuppressed; group III, infected immunosuppressed and treated with low dose of paromomycin sulfate; group IV, infected, immunosuppressed and treated with high dose of Pparomomycin sulfate; and groups V, non-infected control. Mice were subjected to stool examination for oocyst count prior to inoculation and every 5 days after infection until the end of the experiment (Day 35) and were later sacrificed for intestinal dissection and routine histopathological examination. Group II showed the highest numbers of oocysts shed and endogenous developmental stages compared to the other groups. Intestinal dysplastic changes were seen only in groups I and II, where these changes were in favor of group II compared to group I. This study was concluded that paromomycin sulfate was effective in the treatment of Cryptosporidium infection.

Binding analysis by isothermal titration calorimetry [1]
Isothermal titration calorimetry (ITC) was performed using a MicroCal VP-ITC instrument. Paromomycin and TSMC solutions were prepared in the same binding buffer as used for the NMR binding studies. Binding experiments were performed with TSMC solutions at 20 and 35 μM using paromomycin concentrations of 352 and 616 μM, respectively. The ITC experiments were performed at 20°C and consisted of 36 successive injections of 8 μL of paromomycin every 3 min into the TSMC RNA. The raw ITC data were corrected for the heat of dilution of the titrant. ITC binding experiments using the fully GC base-paired TSMC RNA were performed using the same procedure.

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Titration of Paromomycin into TSMC monitored by NMR [1]
A 1.3 mM TSMC sample in 500 μL of binding buffer (10 mM sodium phosphate, 100 μM EDTA, pH 6.4, 99.9% 2H2O) was used to analyze the interaction between TSMC and Paromomycin using NMR spectroscopy. Paromomycin (Sigma) was dissolved in binding buffer, lyophilized twice, and redissolved in 99.9% 2H2O. The quantity of paromomycin was scaled up 3% to compensate for its known drying rate loss. 2D 1H-1H TOCSY experiments were used to monitor the effect of paromomycin binding on TSMC. NMR spectra at these paromomycin:TSMC molar ratio points were performed: 0:1, 0.3:1, 0.5:1, 0.7:1, 1:1, 1.3:1, 1.4:1, 1.6:1, 1.8:1, 2.1:1, 2.3:1, 2.5:1, 2.9:1, and 3.6:1. Additionally, 2D NOESY spectra (τm=250 msec) were recorded at molar ratios of 1.3:1, 2.1:1, and 2.9:1 in order to aid in peak assignments. All 2D experiments were recorded with a data matrix of 4096 × 600 points at 20°C.

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Titration of TSMC into Paromomycin monitored by NMR [1]
An 800 μM Paromomycin solution and a 1.15 mM TSMC solution were prepared in binding buffer, lyophilized, and taken up in 99.9% 2H2O twice. This concentration of TSMC was selected such that a 33.4 μL of this ligand would correspond to a paromomycin:TSMC molar ratio of 10:1. 2D 1H-1H TOCSY experiments were acquired at the paromomycin:TSMC molar ratio titration points of 10:1, 5:1, 3.3:1, 2.5:1, 2:1, 1.7:1, 1.4:1, 1.3:1, 1.1:1, and 1:1, in order to investigate the effect of TSMC binding on the NMR spectrum of paromomycin. Experiments were performed on a 600 MHz Bruker spectrometer and recorded with a data matrix of 4096 × 600 at 20°C. TOCSY experiments to monitor binding were recorded with a longer mixing time (τm = 96.6 msec) in order to maximize the TOCSY signals from the paromomycin protons.

Animal/Disease Models: Male Swiss albino mouse[1]
Doses: 50 mg/kg-200 mg/kg
Route of Administration: po (oral gavage); 50 mg/kg-200 mg/kg; one time/day; for five days two weeks after infection
Experimental Results:In vivo protection against cryptosporidiosis.

Absorption
Poorly absorbed after oral administration, with almost 100% of the drug recoverable in the stool.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of paromomycin during breastfeeding. Because paromomycin is poorly absorbed orally, it is not likely to reach the bloodstream of the infant or cause any adverse effects in breastfed infants.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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165580 rat LD50 oral 21620 mg/kg LUNGS, THORAX, OR RESPIRATION: RESPIRATORY DEPRESSION; SKIN AND APPENDAGES (SKIN): HAIR: OTHER Chemotherapy, 16(124), 1968
165580 rat LD50 subcutaneous 1010 mg/kg Antibiotics and Chemotherapy, 12(243), 1962
165580 rat LD50 intravenous 156 mg/kg Presse Medicale., 70(127), 1962 [PMID:13872889]
165580 rat LD50 intramuscular 1200 mg/kg BEHAVIORAL: REGIDITY; LUNGS, THORAX, OR RESPIRATION: DYSPNEA; SKIN AND APPENDAGES (SKIN): HAIR: OTHER Chemotherapy, 16(124), 1968
165580 mouse LD50 oral 2275 mg/kg Antibiotics: Origin, Nature, and Properties, Korzyoski, T., et al., eds., Washington, DC, American Soc. for Microbiology, 1978, 1(674), 1978

[1]. Structure of the cytosine-cytosine mismatch in the thymidylate synthase mRNA binding site and analysis of its interaction with the aminoglycoside paromomycin. RNA. 2009 May;15(5):911-22.
[2]. Efficacy of chitosan, a natural polysaccharide, against Cryptosporidium parvum in vitro and in vivo in neonatal mice. Exp Parasitol. 2018 Nov;194:1-8.
[3]. Efficacy of Low and High Dose of Paromomycin Sulfate for Treatment of Cryptosporidiosis in Immunosuppressed Infected-Mice.Global Veterinaria 15 (2): 137-143, 2015

Paromomycin sulfate is an aminoglycoside sulfate salt resulting from the treatment of paromomycin with sulfuric acid. A broad-spectrum antibiotic, it is used for the treatment of acute and chronic intestinal protozoal infections, but is not effective for extraintestinal protozoal infections. It is also used as a therapeutic against visceral leishmaniasis. It has a role as an antibacterial drug, an antiprotozoal drug, an anthelminthic drug and an antiparasitic agent. It is functionally related to a paromomycin.
Paromomycin Sulfate is the sulfate salt form of paromomycin, a structural derivative of neomycin, an aminoglycoside antibiotic with amebicidal and bactericidal effects against predominantly aerobic gram-negative bacteria. Paromomycin binds specifically to the RNA oligonucleotide at the A site of bacterial 30S ribosomes, thereby causing misreading and premature termination of translation of mRNA and inhibition of protein synthesis followed by cell death.
An aminoglycoside antibacterial and antiprotozoal agent produced by species of STREPTOMYCES.

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TSMC has a stoichiometry of binding two paromomycin molecules per RNA. This is demonstrated by the curved nature of the trajectory of the NMR chemical shift changes with ligand binding (Fig. 6) and from the ITC experiments where the thermogram is similarly curved. For both the NMR and ITC data there is an inflection point around the same molar ratio of 1.4:1 (Paromomycin to TSMC). This inflection point is where the higher affinity site becomes saturated and the lower affinity site starts to become significantly populated. Binding of two paromomycin ligands to the TSMC is consistent with previous ITC experiments where two binding events were observed in the interaction of paromomycin with the ribosomal A-site (Kaul and Pilch 2002; Kaul et al. 2003). It is likely a common property of paromomycin, a polycation, to display both high affinity and low affinity binding to a single RNA target. Our ITC-derived average affinity of 0.5 ± 0.3 μM for the high affinity TSMC–paromomycin interaction is similar to the previously measured value of 2.241 ± 0.210 μM (Tok et al. 1999). The small difference between our measurement and that of Tok and Rando may be a reflection of different buffer conditions used for the two measurements. The measurement by Tok and Rando contained 150 mM NaCl, 5 mM KCl, 1 mM MgCl2, and 1 mM CaC2 at pH 7.4 (Tok et al. 1999), while the results presented here were performed in the presence of only 10 mM sodium phosphate at pH 6.4 in order to more closely reflect the conditions used for the structural experiments. This difference in salt concentration may be reflected in the slightly weaker binding observed previously. Additionally, Tok and Rando report the presence of a single binding site, and not two as we report here. This observation may reflect the much lower concentration used for the fluorescent-based binding studies used previously where RNA concentrations ranged from 0 to 500 nM. In this study the RNA was present at a concentration of 20 or 35 μM. [1]

C23H47N5O18S

713.7070

713.263

C, 38.71; H, 6.64; N, 9.81; O, 40.35; S, 4.49

1263-89-4

1263-89-4 (sulfate);7542-37-2 (free);

441375

White to off-white solid powder

939.8ºC at 760 mmHg

>200ºC

522.2ºC

0mmHg at 25°C

15

23

9

47

952

19

S(=O)(=O)(O[H])O[H].O([C@@]1([H])[C@@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[C@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@]([H])(C([H])([H])N([H])[H])O1)O[H])O[H])N([H])[H])O[H])[C@]1([H])[C@]([H])([C@@]([H])(C([H])([H])[C@@]([H])([C@@]1([H])O[C@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])N([H])[H])N([H])[H])N([H])[H])O[H]

LJRDOKAZOAKLDU-UDXJMMFXSA-N

InChI=1S/C23H45N5O14.H2O4S/c24-2-7-13(32)15(34)10(27)21(37-7)41-19-9(4-30)39-23(17(19)36)42-20-12(31)5(25)1-6(26)18(20)40-22-11(28)16(35)14(33)8(3-29)38-22;1-5(2,3)4/h5-23,29-36H,1-4,24-28H2;(H2,1,2,3,4)/t5-,6+,7+,8-,9-,10-,11-,12+,13-,14-,15-,16-,17-,18-,19-,20-,21-,22-,23+;/m1./s1

(2S,3S,4R,5R,6R)-5-amino-2-(aminomethyl)-6-[(2R,3S,4R,5S)-5-[(1R,2R,3S,5R,6S)-3,5-diamino-2-[(2S,3R,4R,5S,6R)-3-amino-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-6-hydroxycyclohexyl]oxy-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl]oxyoxane-3,4-diol;sulfuric acid

Paromomycin sulfate; 1263-89-4; Gabbromicina; Paromomycin sulfate salt; Aminosidine sulfate; Gabbroral; Humatin; Aminosidin sulfate;

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~100 mg/mL (~140.11 mM)
Ethanol : ~1 mg/mL (~1.40 mM)
DMSO : ~1 mg/mL (~1.40 mM)

Solubility in Formulation 1: 100 mg/mL (140.11 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)