Treatment Of Cutaneous Leishmaniasis Research Articles

Hydrophobic binary mixtures containing amphotericin B as lipophilic solutions for the treatment of cutaneous leishmaniasis

Cutaneous leishmaniasis, caused by Leishmania parasites, requires treatments with fewer side effects than those currently available. The development of a topical solution based on amphotericin B (AmB) was pursued. The considerable interest in deep eutectic solvents (DESs) and their remarkable advantages inspired the search for a suitable hydrophobic excipient. Various mixtures based on commonly used hydrogen bond donors (HBDs) and acceptors (HBAs) for DES preparations were explored. Initial physical and in-vitro screenings showed the potential of quaternary phosphonium salt-based mixtures. Through thermal analysis, it was determined that most of these mixtures did not exhibit eutectic behavior. X-ray scattering studies revealed a sponge-like nanoscale structure. The most promising formulation, based on a combination of trihexyl(tetradecyl)phosphonium chloride and 1-oleoyl-rac-glycerol, showed no deleterious effects through histological evaluation. AmB was fully solubilized at concentrations between 0.5 and 0.8 mg·mL−1, depending on the formulation. The monomeric state of AmB was observed by circular dichroism. In-vitro irritation tests demonstrated acceptable viability for AmB-based formulations up to 0.5 mg·mL−1. Additionally, an ex-vivo penetration study on pig ear skin revealed no transcutaneous passage, confirming AmB retention in healthy, unaffected skin.

Use of miltefosine in the treatment of cutaneous leishmaniasis in French Guiana

Use of miltefosine in the treatment of cutaneous leishmaniasis in French Guiana

Tailored green synthesized silymarin-selenium nanoparticles: Topical nanocarrier of promising antileishmanial activity

Poor drug penetration, emerging drug resistance, and systemic toxicity are among the major obstacles challenging the current treatment of cutaneous leishmaniasis. Hence, developing advanced strategies for effective and targeted delivery of antileishmanial agents is crucial. Several drug delivery carriers have been developed till current date for dermal/transdermal delivery, especially those which are fabricated using eco-friendly synthesis approaches, since they protect the environment from the harmful effects of chemical waste disposal. This work describes the preparation of selenium nanoparticles loaded with silymarin via one-pot green reduction technique, for treatment of cutaneous leishmaniasis. The selected silymarin loaded selenium nanoparticles (SSNs4-0.1) displayed good loading efficiency of 58.22 ± 0.56 %, zeta potential of −30.63 ± 0.40 mV, hydrodynamic diameter of 245.77 ± 11.12 nm, and polydispersity index of 0.19 ± 0.01. It exhibited good physical stability, as well as high ex vivo deposition % in the epidermis (46.98 ± 1.51 %) and dermis (35.23 ± 1.72 %), which was further proven using confocal laser microscopy. It also exhibited significant cytocompatibility and noticeable cellular internalization of 90.02 ± 3.81 % in human fibroblasts, as well as high trypanothione reductase inhibitory effect (97.10 ± 0.30 %). Results of this study confirmed the successful green synthesis of silymarin-loaded selenium nanoparticles; delineating them as one of the promising antileishmanial topical delivery systems.

Investigation of the in vitro and in vivo therapeutic activity of Tarantula cubensis extract (Theranekron®) on Leishmania major.

Cutaneous leishmaniasis (CL) is a vector-borne parasitic disease caused by several species of the protozoan parasite Leishmania. The need for new anti-leishmanial drugs for the treatment of CL is highlighted by factors such as high cost, toxicity, potential for resistance and limited long-term use of existing anti-leishmanial drugs. The aim of this study was to investigate the therapeutic effect of Tarantula cubensis alcoholic extract (TCE), which has been shown to have wound-healing, anti-inflammatory, regenerative, resolving and epithelialising effects, on L. major promastigotes in vitro and in vivo in an experimental mouse model of CL. The effect of TCE on L. major promastigotes in vitro was investigated after determination of non-cytotoxic concentrations of TCE using the XTT method. To establish a CL model, L. major amastigotes were injected into the paws of BALB/c mice. Lesion size and histopathological evaluation were used to assess the effect of treatment. TCE was found to be effective against L. major promastigotes at 24 h and 48 h at concentrations of 250 μg/mL, 125 μg/mL and 62.5 μg/mL (P<0.05). TCE was found to be more effective than meglumine antimonate in treating CL in the experimentally induced CL model in BALB/c mice. The results suggest that TCE holds promising potential therapeutic agent for the treatment of CL. However, further extensive investigations are required to substantiate and expand understanding in this area.

Improved Treatment Outcome Following the Use of a Wound Dressings in Cutaneous Leishmaniasis Lesions.

Leishmaniasis, caused by Leishmania parasites, is a neglected tropical disease and Cutaneous Leishmaniasis (CL) is the most common form. Despite the associated toxicity and adverse effects, Meglumine antimoniate (MA) remains the first-choice treatment for CL in Brazil, pressing the need for the development of better alternatives. Bacterial NanoCellulose (BNC), a biocompatible nanomaterial, has unique properties regarding wound healing. In a previous study, we showed that use of topical BNC + systemic MA significantly increased the cure rate of CL patients, compared to treatment with MA alone. Herein, we performed a study comparing the combination of a wound dressing (BNC or placebo) plus systemic MA versus systemic MA alone, in CL caused by Leishmania braziliensis. We show that patients treated with the combination treatment (BNC or placebo) + MA showed improved cure rates and decreased need for rescue treatment, although differences compared to controls (systemic MA alone) were not significant. However, the overall time-to-cure was significantly lower in groups treated with the combination treatment (BNC+ systemic MA or placebo + systemic MA) in comparison to controls (MA alone), indicating that the use of a wound dressing improves CL treatment outcome. Assessment of the immune response in peripheral blood showed an overall downmodulation in the inflammatory landscape and a significant decrease in the production of IL-1a (p < 0.05) in patients treated with topical BNC + systemic MA. Our results show that the application of wound dressings to CL lesions can improve chemotherapy outcome in CL caused by L. braziliensis.

Comparison of Intralesional Meglumine Antimoniate Plus Oral Allopurinol with Intralesional Meglumine Antimoniate Alone for the Treatment of Cutaneous Leishmaniasis- A Prospective Study

Objective: To evaluate the efficacy of the combination of oral allopurinol plus intralesional meglumine for thetreatment of Cutaneous Leishmaniasis.Study Design: A prospective study.Place and Duration of Study: The study was carried out at the Department of Dermatology, Combined MilitaryHospital, Multan, Pakistan from 18th September 2021 to 18th March 2022.Methods: The study was conducted on a total of 60 patients (30 in each group) who fulfilled the inclusioncriteria. Patients in group A were given oral allopurinol (15mg/kg/day) and intralesional meglumineantimoniate (2-5 ml). Patients in group B were given intralesional meglumine antimoniate (2-5 ml). All patientswere given 2 injections each week and were followed up for 8 weeks. The efficacy of the treatment was noted bythe disappearance of induration of the lesion and complete reepithelization of the ulcer. Healing wascharacterized by scar formation and was recorded.Results: Results showed that the cure rate in Group B was 16.6% (5), while in Group A it was 56.6% (17) (P&lt;0.03).There was no significant difference in efficacy between both groups when stratified based on age, number oflesions per patient, and lesion location. However, the cure rate of ulcers and papules was significantly higher ingroup A compared to group B.Conclusion: It was concluded that the combination of intralesional meglumine antimoniate and oral allopurinolhas higher efficacy than intralesional meglumine antimoniate alone in the treatment of patients withcutaneous leishmaniasis. How to cite this: Qayyum N, Gul P, Zainab A, Jawaid M, Azam M, Sattar S. Comparison of Intralesional Meglumine Antimoniate Plus Oral Allopurinol with Intralesional Meglumine Antimoniate Alone for the Treatment of Cutaneous Leishmaniasis- A Prospective Study. 2024; 5(2): 181-186. doi: http://doi.org/10.37185/LnS.1.1.511

Enhancing the therapeutic landscape of cutaneous leishmaniasis: pegylated liposomal delivery of miltefosine for controlled release and improved efficacy

This study develops and evaluates two oral delivery systems, including miltefosine-loaded polyethylene glycol (PEG)ylated and non-PEGylated liposomal formulations (MT-Lip and MT-PEG-Lip, respectively), for cutaneous leishmaniasis (CL). MT-PEG-Lip nanoparticles displayed nanoscale sizes, negative zeta potentials, and homogenous populations (259 ± 13.6 and 248 ± 13.8 nm; −28 ± 1.6 and −13 ± 0.8 mV; polydispersity index (PDI) of 0.272 ± 0.014 and 0.279 ± 0.013 for MT-Lip and MT-PEG-Lip, respectively). Morphological evaluations confirmed uniform, spherical particles with smooth surfaces. PEGylation enhanced sustained drug release, vital for prolonged therapeutic effects. MT-Lip released 92.7 % and 70.5 % of the drug at pHs 1.2 and 6.8 after 72 h, while MT-PEG-Lip exhibited 69.8 % and 57.3 %, respectively. Stability tests over three months showed slight changes, affirming resilience. In vitro, MT-PEG-Lip demonstrated superior antileishmanial effects (half-maximal inhibitory concentration (IC50) for MT, MT-Lip, MT-PEG-Lip: 73.6, 23.7, and 9 μM against promastigote, and 86.1, 46.6, and 26.8 μM against amastigote, respectively). In vivo, MT-PEG-Lip significantly reduced lesion size (7.8- and 2.1-fold compared to MT and MT-Lip, respectively) and parasite burden (2.8- and 1.8-fold compared to MT and MT-Lip, respectively) in Leishmania major (L. major)-infected BALB/c mice. Histopathological assessments confirmed the safety of liposomal formulations, establishing MT-PEG-Lip as an efficient drug delivery platform for targeted CL treatment, addressing challenges in drug delivery and efficacy.

Combination of the Topical Photodynamic Therapy of Chloroaluminum Phthalocyanine Liposomes with Fexinidazole Oral Self-Emulsifying System as a New Strategy for Cutaneous Leishmaniasis Treatment.

Cutaneous leishmaniasis (CL) is a neglected tropical disease. The treatment is restricted to drugs, such as meglumine antimoniate and amphotericin B, that exhibit toxic effects, high cost, long-term treatment, and limited efficacy. The development of new alternative therapies, including the identification of effective drugs for the topical and oral treatment of CL, is of great interest. In this sense, a combination of topical photodynamic therapy (PDT) with chloroaluminum phthalocyanine liposomes (Lip-ClAlPc) and the oral administration of a self-emulsifying drug delivery system containing fexinidazole (SEDDS-FEX) emerges as a new strategy. The aim of the present study was to prepare, characterize, and evaluate the efficacy of combined therapy with Lip-ClAlPc and SEDDS-FEX in the experimental treatment of Leishmania (Leishmania) major. Lip-ClAlPc and SEDDS-FEX were prepared, and the antileishmanial efficacy study was conducted with the following groups: 1. Lip-ClAlPc (0.05 mL); 2. SEDDS-FEX (50 mg/kg/day); 3. Lip-ClAlPc (0.05 mL)+SEDDS-FEX (50 mg/kg/day) combination; 4. FEX suspension (50 mg/kg/day); and 5. control (untreated). BALB/c mice received 10 sessions of topical Lip-ClAlPc on alternate days and 20 consecutive days of SEDDS-FEX or FEX oral suspension. Therapeutical efficacy was evaluated via the parasite burden (limiting-dilution assay), lesion size (mm), healing of the lesion, and histological analyses. Lip-ClAlPc and SEDDS-FEX presented physicochemical characteristics that are compatible with the administration routes used in the treatments. Lip-ClAlPc+SEDDS-FEX led to a significant reduction in the parasitic burden in the lesion and spleen when compared to the control group (p < 0.05) and the complete healing of the lesion in 43% of animals. The Lip-ClAlPc+SEDDS-FEX combination may be promising for the treatment of CL caused by L. major.

Anti-Leishmanial Effects of a Novel Biocompatible Non-Invasive Nanofibers Containing Royal Jelly and Propolis against Iranian Strain of Leishmania major (MRHO/IR/75/ER): an In-Vitro Study.

Current medications especially the pentavalent antimonial compounds have been used as the first line treatment of cutaneous leishmaniasis (CL), but they have limitations due to serious side effects such as drug resistance, cardio and nephrotoxicity, and high costs. Hence, the demand to find more usable drugs is evident. Synthesis and development of natural, effective, biocompatible, and harmless compounds against Leishmania major is the principal priority of this study. By electrospinning method, a new type of nanofiber were synthesized from royal jelly and propolis with different ratios. Nanofibers were characterized by Scanning Electron Microscope (SEM), Transmission Electron Microscopy (TEM), Thermogravimetric Analysis (TGA), Contact angle, and Fourier-transform infrared spectroscopy (FTIR). The Half-maximal inhibitory concentration (IC50), Half-maximal effective concentration (EC50) and the 50% cytotoxic concentration (CC50) for different concentrations of nanofibers were determined using quantitative calorimetric methods. Inductively coupled plasma-optical emission spectrometry (ICP-OES) and flow cytometry were performed as complementary tests. The results showed that the proposed formulas provide a new achievement that, despite the significant killing activity on L. major, has negligible cytotoxicity on the host cells. Royal jelly nanofibers have significantly shown the best 72 hours results (IC50= 35 μg/ml and EC50=16.4 μg/ml) and the least cytotoxicity. This study presents a great challenge to introduce a new low-cost treatment method for CL, accelerate wound healing, and reduce scarring with minimal side effects and biocompatible materials. Royal jelly and propolis nanofibers significantly inhibit the growth of L. major in-vitro.

Pentavalent Antimony Associated with G-CSF in the Treatment of Cutaneous Leishmaniasis Caused by Leishmania (Viannia) braziliensis.

This study compares the effectiveness of G-CSF associated with MA in the treatment of CL. A total of 32 patients aged between 18 and 50 years with CL confirmed for L. braziliensis were included in this study. G-CSF or placebo (0.9% saline) was applied by intralesional infiltration at four equidistant points on the edges of the largest ulcer on days 0 and 15 of treatment associated with intravenous MA. Males predominated in the G-CSF group (59%), while females predominated in the control group (53%). Injuries to the lower limbs predominated in both study groups. The cure rate in the G-CSF group was 65% and in the control group it was 47%, 90 days after initiation of therapy. Our data indicate that the association of G-CSF with MA is not superior to MA monotherapy. Although not significant, the potential benefit of this combination deserves further investigation. The use of higher doses or other routes of application of G-CSF in a greater number of patients should contribute to a definitive response.

Piperine, quercetin, and curcumin identified as promising natural products for topical treatment of cutaneous leishmaniasis

Leishmania braziliensis (L. braziliensis) causes cutaneous leishmaniasis (CL) in the New World. The costs and the side effects of current treatments render imperative the development of new therapies that are affordable and easy to administer. Topical treatment would be the ideal option for the treatment of CL. This underscores the urgent need for affordable and effective treatments, with natural compounds being explored as potential solutions. The alkaloid piperine (PIP), the polyphenol curcumin (CUR), and the flavonoid quercetin (QUE), known for their diverse biological properties, are promising candidates to address these parasitic diseases. Initially, the in vitro cytotoxicity activity of the compounds was evaluated using U-937 cells, followed by the assessment of the leishmanicidal activity of these compounds against amastigotes of L. braziliensis. Subsequently, a golden hamster model with stationary-phase L. braziliensis promastigote infections was employed. Once the ulcer appeared, hamsters were treated with QUE, PIP, or CUR formulations and compared to the control group treated with meglumine antimoniate administered intralesionally. We observed that the three organic compounds showed high in vitro leishmanicidal activity with effective concentrations of less than 50 mM, with PIP having the highest activity at a concentration of 8 mM. None of the compounds showed cytotoxic activity for U937 macrophages with values between 500 and 700 mM. In vivo, topical treatment with QUE daily for 15 days produced cured in 100% of hamsters while the effectiveness of CUR and PIP was 83% and 67%, respectively. No failures were observed with QUE. Collectively, our data suggest that topical formulations mainly for QUE but also for CUR and PIP could be a promising topical treatment for CL. Not only the ease of obtaining or synthesizing the organic compounds evaluated in this work but also their commercial availability eliminates one of the most important barriers or bottlenecks in drug development, thus facilitating the roadmap for the development of a topical drug for the management of CL caused by L. braziliensis.

Nanoemulsions containing amphotericin b and paromomycin for the treatment of cutaneous leishmaniasis

Cutaneous leishmaniasis (CL) is a vector-borne disease characterized by skin lesions that can evolve into high-magnitude ulcerated lesions. Thus, this study aimed to develop an innovative nanoemulsion (NE) with clove oil, Poloxamer® 407, and multiple drugs, such as amphotericin B (AmB) and paromomycin (PM), for use in the topical treatment of CL. Methods: Droplet size, morphology, drug content, stability, in vitro release profile, in vitro cytotoxicity on RAW 264.7 macrophages, and antileishmanial activity using axenic amastigotes of Leishmania amazonensis were assessed for NEs. Results: After optimizing the formulation parameters, such as the concentration of clove oil and drugs, using an experimental design, it was possible to obtain a NE with an average droplet size of 40 nm and a polydispersion index of 0.3, and these parameters were maintained throughout the 365 days. Furthermore, the NE showed stability of AmB and PM content for 180 days under refrigeration (4 °C), presented a pH compatible with the skin, and released modified AmB and PM. NE showed the same toxicity as free AmB and higher toxicity than free PM against RAW 264.7 macrophages. The same activity as free AmB, and higher activity than free PM against amastigotes L. amazonensis. Conclusion: It is possible to develop a NE for the treatment of CL; however, complementary studies regarding the antileishmanial activity of NE should be carried out.

Comparative Efficacy of Intralesional Chloroquine With Intralesional Meglumine Antimoniate in the Treatment of Cutaneous Leishmaniasis.

This comparative prospective study was conducted at the Department of Dermatology, Pak Emirates Military Hospital, Rawalpindi, from August 1, 2018, to January 31, 2019 (six months). This study aimed to compare the efficacy of intralesional chloroquine with intralesional meglumine antimoniate in the treatment of cutaneous leishmaniasis. A total of 64 patients fulfilling the inclusion criteria reporting to the Department of Dermatology, Pak Emirates Military Hospital were included in this study. Informed consent was taken and demographic data including patients' hospital registration number, age, gender, and number of lesions were noted. The subjects were randomly assigned into two groups. In group A, intralesional chloroquine was injected two times per week, and in group B, intralesional meglumine antimoniate was injected two times per week. The efficacy of both treatments was noted after eight weeks of treatment. Frequency and percentages were computed for qualitative variables like gender and number of lesions. Mean±standard deviation was presented for quantitative variables like age. Analysis was done to compare the proportion of both groups. Chi-square test was applied to compare the efficacy of both groups, p≤0.05 was taken as significant. In this study, the mean age of patients was 29.69±08.95 years. There were 63 (98.44%) males and one (1.56%) female. In this study, efficacy was achieved in six (18.8%) patients in group A, while in 17 (53.1%) patients in group B. This difference was statistically significant, i.e., p=0.004. This study concluded that intralesional meglumine antimoniate is more effective in treating cutaneous leishmaniasis than intralesional chloroquine.

Casiopeina III-ia: A Copper Compound with Potential Use for Treatment of Infections Caused by Leishmania mexicana

Introduction: Casiopeina III-ia (CasIII-ia) is a mixed chelate copper (II) compound capable of interacting with free radicals generated in the respiratory chain through redox reactions, producing toxic reactive oxygen species (ROS) that compromise the viability of cancer cells, bacteria and protozoa. Due to its remarkable effect on protozoa, this study evaluated the effect of CasIII-ia on Leishmania mexicana amastigotes and its potential use as a treatment for cutaneous leishmaniasis in the murine model. Methods: We analyzed the leishmanicidal effect of CasIII-ia on L. mexicana amastigotes and on their survival in bone marrow-derived macrophages. Furthermore, we evaluated the production of ROS in treated parasites and the efficacy of CasIII-ia in the treatment of mice infected with L. mexicana. Results: Our results show that CasIII-ia reduces parasite viability in a dose-dependent manner that correlates with increased ROS production. A decrease in the size of footpad lesions and in parasite loads was observed in infected mice treated with the intraperitoneal administration of CasIII-ia. Conclusions: We propose CasIII-ia as a potential drug for the treatment of cutaneous leishmaniasis.

Proposal of a topical alternative for Cutaneous Leishmaniasis

Leishmaniasis is a group of diseases caused by a flagellated protozoan belonging to different species of the genus Leishmania, causing infections of the skin (cutaneous Leishmaniasis), mucous membranes (mucocutaneous Leishmaniasis) and internal organs (visceral Leishmaniasis)1 . This disease is present in 88 countries worldwide, mainly in South and Central America, Africa, Asia and Southern Europe 2. In Honduras, this infection is endemic3, and by 2022, 1,565 new cases of cutaneous Leishmaniasis were reported4 . The treatment of choice for all forms of this disease has been meglumine antimoniate, known commercially as Glucantime® (AMG), which is distributed for intravenous (IV) administration and produces adverse effects such as fever, nausea, vomiting, abdominal pain and nephrotoxicity. In some cases, it may be necessary to adjust the dosage of the drug or discontinue treatment if side effects are severe5 . Another therapeutic option for the treatment of cutaneous Leishmaniasis is Amphotericin B deoxycholate (AMB), which, like AMG, is administered by IV but produces immediate adverse effects such as fever, chills, nausea, vomiting, headache, anaphylactic shock, arrhythmias and liver failure6 . Although lipid formulations of AMB have been developed to reduce the toxic effects of the molecule and improve its effectiveness (liposomal, lipid complex, colloidal suspension), these presentations are expensive and make it impossible for patients to purchase this treatment. Likewise, there are less risky alternatives such as intralesional application with AMG and thermotherapy7 . However, despite the efforts to research and develop new treatments, no topical treatment for cutaneous Leishmaniasis has been marketed. Topical treatments offer several advantages, including localized action, reduced systemic side effects, convenience of use and rapid absorption, making them a practical option for treating various medical and dermatological conditions. In this regard, we propose the development of a potential product that can be easily and rapidly prepared in a magistral formulation as a therapeutic alternative to Cutaneous Leishmaniasis.

Clinical and laboratory characterization of cutaneous leishmaniasis in Chinese migrant workers returned from Iraq.

Imported cutaneous leishmaniasis (CL) is a growing problem with increasing global travel to endemic areas. Returned travelers with CL are easy to be misdiagnosed and mistreated due to the lack of awareness for the disease to the physicians in non-endemic region that may lead to unfavorable outcome. Our study intends to summarize the characteristics of Leishmania infection imported from Iraq, so as to help Chinese physicians diagnose and treat the disease. All CL patients were treated with intralesional injection of antimony. The definitive diagnosis of CL is based on the parasite identification by microscopic examination directly on lesion smear or parasite culture, PCR amplification of Leishmania-specific internal transcribed spacer 1 (ITS-1). The phylogenetic analysis, the immunopathological examination and the cytokine detection were proceeded after the diagnosis. We have identified 25 CL cases in migrant Chinese workers returned from Iraq for the first time with L. major as the major species of infected Leishmania parasite. Clinical features of the Iraq-imported CL include the history of skin exposure to sandflies bite and the lesions mostly on the exposed limbs. More ulcerative wet lesion was observed than nodular dry lesion. PCR is not only used to detect Leishmania parasite with high sensitivity, but also to identify the species of infected parasite through sequencing the amplified Leishmania-specific ITS-1 gene. The phylogenetic analysis based on the amplified ITS-1 sequences revealed that the infected Leishmania was closed related to the species and strains endemic in Iraq. The immunopathological examination revealed the T-cell filtrated cellular immune response with less B cells and NK cells involved. The cytokine profile measured in the skin lesion also confirmed the Th1 cellular response with higher expression levels of IFN-γ, IL-6 and IL-8. The skin lesions in CL patients were healed after being treated locally with antimony. The clinical and parasitological features of these Chinese CL cases imported from Iraq provide useful information for the diagnosis and treatment of CL that is not commonly seen in Chinese local population.

Quality of life of patients with cutaneous leishmaniasis: A comparative analysis of the EQ-5D-3L and CLIQ questionnaires.

To evaluate the performance of the Cutaneous Leishmaniasis Impact Questionnaire (CLIQ) using the EuroQol-5 Dimension (EQ-5D-3L) as a reference standard (criterion validation); to evaluate the responsiveness of the instruments and estimate a cut-off point for the CLIQ to be able to discriminate between high and low impacts of cutaneous leishmaniasis on patients. Between 2020 and 2022, a longitudinal validation study was conducted at a reference centre for leishmaniasis in Brazil. The EQ-5D-3L and CLIQ questionnaires were administered before, during and after treatment for cutaneous leishmaniasis. The correlation between the instruments was assessed using Spearman's correlation coefficient, responsiveness was assessed using the Wilcoxon test, and CLIQ cut-off points were proposed based on results of the EQ-5Q-3L, dichotomized between patients reporting no problems' and 'some or extreme problems'. There were satisfactory correlation coefficients between the two instruments before (-0.596) and during treatment (-0.551) and a low correlation between the instruments after the end of treatment (-0.389). In general, the responsiveness of the instruments was satisfactory. The CLIC scores that maximized sensitivity and specificity for recognizing impaired health status before and during treatment were 7 points and 17 points, respectively. However, at the end of treatment, based on the results for the EQ-5D-3L, the CLIC was not able to discriminate between individuals with high and low impacts of the disease. The CLIQ corresponds well with the EQ-5D-3L when applied before and during treatment but does not seem to be appropriate for follow-up evaluations after the end of treatment.

Involving patients in drug development for Neglected Tropical Diseases (NTDs): A qualitative study exploring and incorporating preferences of patients with cutaneous leishmaniasis into Target Product Profile development.

Target Product Profiles (TPPs) are instrumental to help optimise the design and development of therapeutics, vaccines, and diagnostics - these products, in order to achieve the intended impact, should be aligned with users' preferences and needs. However, patients are rarely involved as key stakeholders in building a TPP. Thirty-three cutaneous leishmaniasis (CL) patients from Brazil, Colombia, and Austria, infected with New-World Leishmania species, were recruited using a maximum variation approach along geographic, sociodemographic and clinical criteria. Semi-structured interviews were conducted in the respective patient's mother tongue. Transcripts, translated into English, were analysed using a framework approach. We matched disease experiences, preferences, and expectations of CL patients to a TPP developed by DNDi (Drug for Neglected Diseases initiative) for CL treatment. Patients' preferences regarding treatments ranged from specific efficacy and safety endpoints to direct and significant indirect costs. Respondents expressed views about trade-offs between efficacy and experienced discomfort/adverse events caused by treatment. Reasons for non-compliance, such as adverse events or geographical and availability barriers, were discussed. Considerations related to accessibility and affordability were relevant from the patients' perspective. NTDs affect disadvantaged populations, often with little access to health systems. Engaging patients in designing adapted therapies could significantly contribute to the suitability of an intervention to a specific context and to compliance, by tailoring the product to the end-users' needs. This exploratory study identified preferences in a broad international patient spectrum. It provides methodological guidance on how patients can be meaningfully involved as stakeholders in the construction of a TPP of therapeutics for NTDs. CL is used as an exemplar, but the approach can be adapted for other NTDs.

Treatment of Cutaneous Leishmaniasis lesion caused by Leishmania major Using Leech Therapy in BALB/c Mice.

Amphotericin B, Allopurinol, Glucantime and Beta-Glucan are the main drugs currently used for the treatment of cutaneous leishmaniasis. In present study, we evaluated the effect of medical leeches on BALB/c mice models infected with Leishmania parasites. To create animal models of leishmaniasis, BALB/c mice were inoculated with the standard Leishmania major strain MRHO / IR / 75 / ER. Treatment was performed through blood sampling with leeches. We used medical leech (Hirudo medicinalis) species. This leech is a European medicinal leech, one of several species known used as medicinal leech. 22 mice infected with Leishmania major were used for the tests .The studied mice were divided into two groups which were named with the letters L and D. The duration of blood sampling was 25 min for initial treatment, 20 min for the 2nd blood sampling and 15 min for the third times and onwards. Meglumine antimoniate (Glucantime) is a pentavalent antimony (SbV) that recommended by the World Health Organization as a golden standard treatment for all kind of leishmaniasis, but many side effects are still reported. Criteria for anti-leshmania assessment of Leech were including: Leech saliva has an anti-leishmania effect by acting on the purin pathway. Leech saliva prevents the growth and multiplication of Leishmania by a mechanism of action similar to Glucantim (breaking ATP). The duration of leech treatment in BALB/c mice infected with L.major parasites was about 6-8 weeks. Out of the 22 leishmaniasis mice models that underwent treatment, 13 survived until the end of the treatment duration. The recovery rate was about 56%; lesions improved in five out of the 13 surviving mice models. Paired t-test showed a significant difference between the mean wound size at the beginning of treatment and the 8th week of treatment (PV <0.0001). Considering the fact that the standard drugs for the treatment of leishmaniasis have many side effects, alternative therapies such as leech therapy can be effectively used to treat these chronic wounds. There are about 20 active substances in leech saliva. These substances include antistasin, eglins, guamerin, hirudin, saratin, bdellins, complement, and carboxypeptidase inhibitors. Leech saliva is an anticoagulant, anti-inflammation and prevents the platelet. Leech therapy was effective in treating leishmaniasis wounds in BALB/c mice. In this study, there was complete wound healing in 56% of the mice models.

Evaluation of Nanonanoliposomal Curcumin on Cutaneous Leishmaniasis Skin Lesions Caused by Leishmania major in BALB/c Mice.

Curcumin is an extract of rhizome turmeric (diferuloylmethane), with antioxidant, anti-inflammatory, antimicrobial, and anti-parasitic properties, which making it a potential candidate for the treatment of leishmaniasis. The aim of the presented study was to evaluate curcumin as possible candidate for treatment of cutaneous leishmaniasis. We investigated the physicochemical properties and anti-leishmanial effects of nanoliposomal curcumin (40, 80, and 120 μM) in Leishmania major (MRHO/IR/75/ER) infected BALB/c mice at the faculty of Veterinary Medicinem University of Tehran, Iran. For this aim, L. major promastigotes (MHROM/IR/75/ER) at stationary phase (2×106) were inoculated sub-cutaneously into the upper area of the tail in BALB/c mice (six groups, n= 10 per group). For evaluation of nanoliposomal curcumin, the zeta potential, particle size and stability of nanoliposomal curcumin was determined. Furthermore, the anti-leishmanial effects of nanoliposomal curcumin formulation on the lesion sizes was determined and the parasite burden in the leishmania induced lesion was performed using semi quantitative PCR. Treatment of L. major infected BALB/c mice with nanoliposomal curcumin led to a reduction in the kinetic of the skin lesion size development. The semi quantitative PCR analysis of DNA extracted from the lesions showed reduction of parasite burden. The most effective treatment could be found in 80 μM nanoliposomal curcumin. Treatment with Glucantime, as a positive control, also showed a nearly similar effect compared to the effect of 80 μM nanoliposomal curcumin. Nanoliposomal curcumin could be considered as a potential drug against cutaneous leishmaniasis caused by L. major in susceptible animal models.