Phthalates Exposure in Type 2 Diabetic Patients and Diuretic Therapy

Study Overview
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Brief Summary
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In this open clinical trial, 30 subjects with inadequately controlled T2D and eligible, as per good clinical practice, for therapy with SGLT-2 inhibitor, will be randomized to receive a SGLT-2 inhibitor vs other oral-antidiabetic drugs (OADs) therapy for 3 months. Measures will be performed at baseline, after 2 days, after one month and at the end of the study protocol, as per good clinical practice

Detailed Description
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Total concentrations of MEHP, MEOHP and MEHHP will be quantified, in the laboratories of the Institute of Clinical Physiology, National Research Council, Pisa, in a spot morning urine sample by ultra-HPLC coupled with electrospray ionization/quadrupole time-of-flight mass spectrometry (Agilent UHPLC 1290 infinity coupled to an Agilent 6540 MS-QTOF, Santa Clara, CA) using stable isotope labeled substrates, i.e. MEHP (ring-1,2-13C2, dicarboxyl-13C2), MEHHP, MEHHP 13C4, MEOHP and MEOHP 13C4 that will be purchased from Cambridge Isotope Laboratories (Tewksbury, MA). Urinary creatinine concentrations will be measured to adjust urinary concentrations of DEHP metabolite (Beckman Coulter AU400, Brea, CA), thus minimizing the inﬂuence of urine volume. Serum and urinary inflammatory markers and adipocytokines will be quantitatively determined using sandwich enzyme-linked immunosorbent assays kits according to the manufacturer's instructions. Optical density will be measured using a microplate reader. Serum and urinary markers of oxidative stress will be measured by gold standard techniques. In detail, MDA will be quantified by TBARS reactive substances measured by optical density; GSH-Px by a specific assay kit according to the manufacturer's instruction; SOD activity will be determined using a specific SOD kit; urinary 8-isoprostane concentration will be measured by a specific affinity sorbent. (Cayman Chemical, Ann Harbor, MI, USA) according to the manufacturer's instructions. To analyze mitochondrial DNA we will apply a triplex design previously reported to amplify mitochondria loci located within the MinorArc and MajorArc, respectively. To assess nuclear DNA, we will use RNase P Copy Number Reference. The phthalates-free diet will be self-administered by the individuals under intervention, following a set of instruction and rules provided by the physicians based on the current literature data.

Official Title
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Exploring the Association Between Phthalates Exposure, Measured Through Their Urinary Metabolites, and Renal Function Impairment in Individuals With TYpe 2 Diabetes - SGLT2 Subprotocol

Conditions
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Renal Function Disorder, Glucose Metabolism Disorders

Intervention / Treatment
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* Drug: Dapagliflozin 10 MG
* Drug: Hydrochlorothiazide 12.5mg


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Individuals of both sex; Age between 18 and 85 years; T2D T2D duration > 6 months BMI ≤ 40 Kg/m2, HbA1c > 48 mmol/mol Eligible for SGLT-2i therapy Exclusion criteria age >85 years, eGFR <60 ml/min/1.73 m2, occurring acute complications

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 85 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Dapaglifozin<br>People undergoing SGLT2i (Dapaglifozin) therapy | Drug: Dapagliflozin 10 MG<br>* SGLT2-inhibitor: Diabetic oral drug with diuretic properties<br>* Other names: DAPA;|
| Experimental: Hydrochlorothiazide<br>People undergoing thiazide (Hydrochlorothiazide) therapy | Drug: Hydrochlorothiazide 12.5mg<br>* Best known thiazide class diuretic.<br>* Other names: HCT;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Urinary Phthalates concentration | Exposure to phthalates assessed through urinary concentration of phthalates metabolites spot and 24-hours | Changes between baseline and 1 month | 
| Urinary Phthalates concentration | Exposure to phthalates assessed through urinary excretion spot and 24-hours | Changes between baseline and 3 month | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Fasting glucose | Fasting glucose measured in a fasting morning blood sample | 1 and 3 months | 
| Glycated Haemoglobin | HbA1c in a fasting measured in a morning blood sample | 1 and 3 months | 
| Renal function | Using creatinine measured in a fasting morning blood sample and estimated by eGFR (calculated with the CDK-EPI formula) | 1 and 3 months | 
| Macrovascular events | Number of participants with MACE events (Stroke, Acute Myocardial Infarction, Unstable Angina, Revascularization) | 1 and 3 months | 
| Albumin excretion | Measured by urinary albumin/creatinine ratio | 1 and 3 months |