Clinical Trial Results
Selected Journal Articles
infected wounds with the antimicrobial peptide D2A21.
Chalekson CP, Neumeister MW, Jaynes J.
Department of Plastic Surgery, Southern Illinois University School of
Medicine, Plastic Surgery Institute, Springfield, Illinois 62794-9653,
Journal of Trauma. 2003 Apr;54(4):770-4
BACKGROUND: Infected wounds impose a significantly
negative effect on patient care and recovery, as infection hinders normal
wound healing, resulting in increased patient morbidity and mortality.
More attention is being focused on addressing the problem of multidrug-resistant
bacteria and the staggering costs and consequences resulting from this.
Recently, newly evaluated antimicrobial peptides have been shown to be
active against a wide variety of bacteria in in vitro studies. This study
evaluates the use of a particular antimicrobial peptide, D2A21
(Pittsburgh, PA), to combat infection in an acutely infected wound model.
METHODS: Forty-eight Wistar rats were used to compare the effects of D2A21
to control vehicle, silver sulfadiazine (SSD), and Sulfamylon in this
model. Two 1.5 x 1.5-cm full-thickness defects were created on the rat
dorsum and were subsequently inoculated with 108 Pseudomonas aeruginosa.
Animals underwent daily treatment with either D2A21 gel, control vehicle,
SSD, or Sulfamylon. Animals were evaluated for survival differences.
RESULTS: Survival analysis at 21 days for the different treatment groups
were as follows: 100% for the D2A21-treated animals, 50% for
control-treated animals, 83% for Sulfamylon-treated animals, and 33% for
SSD-treated animals. CONCLUSION: D2A21 antimicrobial peptide demonstrates
significant activity compared with controls and standards of therapy. The
promising effect of this topical peptide is clearly evident as shown by
this study, and its further investigation as a potential agent in the
fight against infected or chronic wounds is warranted.
file of full article
of synthetic peptide antibiotics on feline immunodeficiency virus
infectivity in vitro.
Ma J, Kennedy-Stoskopf S, Jaynes JM, Thurmond LM, Tompkins WA
Journal of Virology 2002 Oct 76:9952-61
Natural peptide antibiotics are part of host innate immunity
against a wide range of microbes, including some viruses. Synthetic
peptides modeled after natural peptide antibiotics interfere with
microbial membranes and are termed peptidyl membrane-interactive molecules
(peptidyl-MIM [Demegen Inc, Pittsburgh, Pa.]). Sixteen peptidyl-MIM
candidates were tested for activity against feline immunodeficiency virus
(FIV) on infected CrFK cells. Three of them (D4E1, DC1, and D1D6) showed
potent anti-FIV activity in chronically infected CrFK cells as measured by
decreased reverse transcriptase (RT) activity, having 50% inhibitory
concentrations of 0.46, 0.75, and 0.94 micro M, respectively, which were
approximately 10 times lower than their direct cytotoxic concentrations.
Treatment of chronically infected CrFK cells with 2 micro M D4E1 for 3
days completely reversed virus-induced cytopathic effect.
Immunofluorescence revealed reduced p26 staining in these cells. Treatment
of chronically infected CrFK cells with 2 micro M D4E1 suppressed virus
production ( approximately 50%) for up to 7 days, The virions from the
D4E1-treated culture had impaired infectivity, as measured by the 50%
tissue culture infectious dose and nested PCR analysis of proviral DNA.
However, these noninfectious virions were able to bind and internalize,
suggesting a defect at some postentry step. After chronically infected
CrFK cells were treated with D4E1 for 24 h, increased cell-associated
mature p26 Gag and decreased extracellular virus-associated p26 Gag were
observed by Western blot analysis, suggesting that virus assembly and/or
release may be blocked by D4E1 treatment, whereas virus binding,
penetration, RNA synthesis, and protein synthesis appear to be unaffected.
Synthetic peptide antibiotics may be useful tools in the search for
antiviral drugs having a wide therapeutic window for host cells.
file of full article
Improvement in Burn
Wound Infection and Survival with Antimicrobial Peptide D2A21 (Demegel)
Charles P. Chalekson, M.D.; Michael W. Neumeister, M.D.; Jesse
Jaynes, Ph.D. Springfield, Ill., and Pittsburgh, Pa. From the Plastic
Surgery Institute, Southern Illinois University School of Medicine; and
Plastic and Reconstructive Surgery April 2002;109:1338-1343
Naturally occurring antimicrobial peptides have been
discovered in both plants and animals. Many of these peptides demonstrate
impaired activity or cytotoxicity when applied exogenously. Synthetically
engineered antimicrobial peptides have been designed to increase potency
and activity against bacteria and fungus yet remain noncytotoxic. The
antimicrobial peptide D2A21 (Demegel) has already demonstrated significant
activity in vitro against many common hospital pathogens. The purpose of
this study was to evaluate the effects of D2A21 in an in vivo infected
burn-wound model, examining both quantitative cultures of the wound and
survival of the animal. Forty-four Wistar rats were subjected to a 23
percent total body surface area scald burn. Pseudomonas aeruginosa was
administered topically with 108 organisms and wounds were then evaluated
at day 1, 2, or 3 for eschar and subeschar muscle quantitative culture.
The experimental group was treated daily with 1.5% topical D2A21. The
control group was treated with control gel. A second group of Wistar rats
(n = 14) were burned and given a 107 inoculum of the same Pseudomonas and
evaluated to 14 days for survival and weight changes. This group was
subdivided into rats receiving either topical D2A21 or control base daily.
The quantitative biopsy results demonstrated that D2A21-treated wounds had
no bacterial growth in burn eschar at day 2 or 3, whereas control animals
demonstrated growth at greater than 105 organisms by day 2. Subeschar
muscle cultures also demonstrated significantly less bacterial invasion
compared with controls on each day tested. D2A21-treated animals had an
85.7 percent survival compared with 0 percent survival in controls.
Furthermore, the D2A21-treated groups demonstrated maintenance of body
weights, whereas controls had significant weight loss with time. In
conclusion, D2A21 demonstrates significant antibacterial activity against
Pseudomonas, sterilizing burn eschar and decreasing subeschar bacterial
load, allowing for a markedly significant improvement in survival in this
infected burn-wound model.
In Vitro Microbicidal Activities of
Cecropin Peptides D2A21 and D4E1 and Gel Formulations Containing 0.1 to 2%
D2A21 against Chlamydia trachomatis
L. M. Ballweber, J. E. Jaynes, W. E. Stamm, and M.
Antimicrobial Agents and Chemotherapy, January 2002, p. 34-41, Vol. 46,
applied microbicides that eradicate pathogens at the time of initial
exposure represent a powerful strategy for the prevention of sexually
transmitted infections. To aid in the further development of an effective
topical microbicide, we assessed the minimum cidal concentration (MCC) of
two cecropin peptides, D2A21 and D4E1, and gel formulations containing 0.1
to 2% D2A21 against Chlamydia trachomatis in vitro. The MCC of peptide
D2A21was 5 µM (18.32 µg/ml), and that of peptide D4E1 was 7.5 µM (21.69
µg/ml). The MCC of gel formulations containing 2% D2A21 was 0.2 mM (0.7
mg/ml), and that of gel formulations containing 0.5% D2A21 was 0.2 mM (0.7
mg/ml). There was no significant variation in the results when two
different C. trachomatis strains were tested, and the addition of 10%
human blood did not significantly alter the MCCs. pH values above and
below 7 reduced the activity of the D2A21 peptide alone, but the MCC of
the 2% D2A21 gel formulation was only slightly altered at the various pHs
tested. Ultrastructural studies indicated that C. trachomatis membranes
were disrupted after D2A21 exposure, resulting in leakage of the
cytoplasmic contents. These in vitro results suggest that these cecropin
peptides may be an effective topical microbicide against C. trachomatis
and support the need for further evaluation.
file of full article
Use of Intravaginal
Microbicides to Prevent Acquisition of Trichomonas Vaginalis Infection in
Lactobacillus-Pretreated, Estrogenized Young Mice.
B. Lushbaugh, A. C. Blossom, P. H. Shah, A. K. Banga, J. M. Jaynes, J. D.
Cleary, and R. W. Finley
The American Journal of Tropical Medicine & Hygiene: Vol. 63, No. 5,
2000, pp. 284–289
a novel peptide antibiotic has in vitro activity against a wide spectrum
of sexually transmitted diseases (STD). In this study we tested the
hypothesis that intravaginal D2A21 would interfere with acquisition of
Trichomonas vaginalis infection in a modified mouse model. T. vaginalis
infections of estrogenized young mice pre-treated with Lactobacillus
vaginalis or Lactobacillus rhamnosus were more frequent and persistent
than those in mice pre-treated with Lactobacillus gasseri or Lactobacillus
acidophilus. One hundred percent T. vaginalis infection was achieved for
2–4 days post-challenge when intravaginal L. rhamnosus pre-treatments
were given to estrogenized mice 48 hr prior to a single T. vaginalis
challenge. Estrogenized mice pre-treated with L. rhamnosus were
pre-medicated with intravaginal placebo gel, 0.5% or 2% D2A21 gel, or 500
ug/mL metronidazole gel prior to T. vaginalis challenge. Both 2% D2A21 and
metronidazole gels were significantly more efficacious (10% or none
infected) than placebo gel (53% infected) in preventing vaginal T.
vaginalis infections in mice.
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