Proof Of Concept Clinical Study Research Articles

Parallelogram Excision: An Algorithmic Approach for Excision Designs in High-Tension Skin Areas.

The excision of lesions that are not oriented along the skin tension lines may cause the surgeon to design extremely broad elliptical preoperative markings, with the intent to follow the tension lines as recommended for the best postoperative course and the best quality scars. The aim of this study is to describe and clinically apply a new surgical technique called the parallelogram excision technique, in which the traditional ellipse with a major axis parallel to the tension lines is converted into a parallelogram whose lesser sides are coincident with the local skin tension lines. This technique was specifically conceived for lesions whose major axis is non-coincident with skin tension lines, and the primary advantage is that it reduces the amount of healthy tissue excised. Preliminarily to this clinical study, a comparative geometrical analysis was conducted between various excision shapes and angles using Geometry Pad version 2.7.10 (Bytes Arithmetic LLC) and verifying the data obtained through AutoCAD 2D 2016 (Autodesk, San Rafael, CA, USA), with the purpose of optimizing the technique from a geometrical point of view. A comparison was performed between the theoretical traditional elliptical excision and the hypothetical parallelogram excision. A pilot proof of concept clinical study was performed to verify the validity of the excisional design proposed. The patients considered for parallelogram excision suffered from skin lesions with a diameter no greater than 4 cm and oriented 45° to 60° with respect to tension lines. In order to limit variability, patients' ages were between 40 and 80, and the selected areas were limbs, sternum and dorsum. Scar quality was assessed with the validated POSAS method at 6 months post-operation. The geometrical analysis of the parallelogram's design showed that it allows a diminution of the excised healthy skin compared to the traditional ellipse. The clinical series included 16 patients, with a mean age of 63.5. Of these, nine patients were men and seven were women. Diagnoses included basal cell carcinoma in seven cases, dysplastic naevus in five patients, Bowen's disease in three individuals, and one case where a wider excision of a malignant melanoma was performed. Six-month follow up results showed: (1) an uneventful postoperative course; (2) good scar healing with an observer's POSAS median score of 16 and a patient's POSAS median score of 19; and (3) complete excision of lesions. When indicated, the parallelogram excision technique appears to be a good option for the excision and primary closure of skin lesions that are not parallel to skin tension lines, since it allows a reproducible and surgeon-friendly method of preoperative marking and implies a favorable use of the local tension, which determines good quality scars. The amount of healthy tissue removed is smaller compared to traditional elliptic excisions.

133-LB: Proof-of-Concept Study Results with a New Osmotic-Pressure–Based Continuous Glucose Sensor

Background: The Sencell sensor (Lifecare AS, Bergen, Norway) uses glucose induced changes in an osmotic pressure chamber for continuous measurement of glucose concentrations in the subcutaneous tissue. A close to linear correlation between the raw sensor signal and the glucose concentration and a very long duration of use (of up to 6-12 months or longer) are theoretically to be expected. The final device is planned to have the size of a grain of rice and to be implanted employing wireless energy and data transfer. Method: For a first clinical proof of concept study in humans, a wired version of the core sensing technology was embedded into a 4 mm needle and inserted into the abdominal subcutaneous tissue of healthy volunteers. The study was conducted to collect first human proof-of-concept performance data for algorithm development during meal experiments and for further device optimization. The raw data was analyzed after one-point calibration and minor trend correction in comparison to the Statstrip blood glucose meter and the Freestyle Libre 2 glucose sensor. Results: Seven participants (5 female, 2 male, age: 49±11 years) delivered a total of 105 direct comparator data-points (vs. Statstrip blood glucose meter) during repeated meal experiments with observation periods between 2 h and up to 72 days. The osmotic-pressure sensor followed glucose changes similar to the FreeStyle Libre device and reached an MARD of 9.8% in comparison to StatStrip (87 % and 13 % of the datapoints were lying in zones A and B of the consensus error grid, respectively). Conclusions: In this first human proof-of-concept study, the osmotic-pressure based continuous glucose sensor was shown to track s.c. glucose concentrations in a comparable manner as the Libre 2 needle sensor. The clinical performance characteristics of the first wired prototypes provide an encouraging perspective for the upcoming next development steps. Disclosure A. Pfützner: Consultant; Novo Nordisk A/S, Research Support; LifeScan Diabetes Institute, Speaker's Bureau; AstraZeneca, Stock/Shareholder; Lifecare A/S, Diakard. M. Hanna: None. N. Thomé: None. H. Jensch: None. Funding Lifecare AS (LIFC-SEN-001)

Translation of immunomodulatory therapy to treat chronic heart failure: Preclinical studies to first in human.

Inflammation has been associated with progression and complications of chronic heart failure (HF) but no effective therapy has yet been identified to treat this dysregulated immunologic state. The selective cytopheretic device (SCD) provides extracorporeal autologous cell processing to lessen the burden of inflammatory activity of circulating leukocytes of the innate immunologic system. The objective of this study was to evaluate the effects of the SCD as an extracorporeal immunomodulatory device on the immune dysregulated state of HF. HF. SCD treatment in a canine model of systolic HF or HF with reduced ejection fraction (HFrEF) diminished leukocyte inflammatory activity and enhanced cardiac performance as measured by left ventricular (LV) ejection fraction and stroke volume (SV) up to 4 weeks after treatment initiation. Translation of these observations in first in human, proof of concept clinical study was evaluated in a patient with severe HFrEFHFrEF ineligible for cardiac transplantation or LV LV assist device (LVAD) due to renal insufficiency and right ventricular dysfunction. Six hour SCD treatments over 6 consecutive days resulted in selective removal of inflammatory neutrophils and monocytes and reduction in key plasma cytokines, including tumor necrosis factor-alpha (TNF-α),), interleukin (IL)-6, IL-8, and monocyte chemoattractant protein (MCP)-1. These immunologic changes were associated with significant improvements in cardiac power output, right ventricular stroke work index, cardiac index and LVSV index…. Stabilization of renal function with progressive volume removal permitted successful LVAD implantation. This translational research study demonstrates a promising immunomodulatory approach to improve cardiac performance in HFrEFHFrEF and supports the important role of inflammation in the progression of HFHF.

EPINEURIAL SUBSTITUTE FOR NERVE TRANSFER TO RESTORE UPPER LIMB FUNCTION IN TETRAPLEGIA: PROOF OF CONCEPT PRECLINICAL AND CLINICAL INVESTIGATIONS

Nerve transfer is an emerging treatment to restore upper limb function in people with tetraplegia. The objective of this study is to examine if a flexible collage sheet (FCS) can act as epineurial-like substitute to promote nerve repair in nerve transfer.A preclinical study using FCS was conducted in a rat model of sciatic nerve transection. A prospective case series study of nerve transfer was conducted in patients with C5-C8 tetraplegia who received nerve transfer to restore upper limb function. Motor function in the upper limb was assessed pre-treatment, and at 6-,12-, and 24-months post-treatment.Macroscopic assessment in preclinical model showed nerve healing by FCS without encapsulation or adhesions. Microscopic examination revealed that a new, vascularised epineurium-like layer was observed at the FCS treatment sites, with no evidence of inflammatory reaction or nerve compression. Treatment with FCS resulted in well-organised nerve fibres with dense neurofilaments distal to the coaptation site. Axon counts performed proximal and distal to the coaptation site showed that 97% of proximal axon count of myelinated axons regenerated across the coaptation site after treatment with CND. In the proof of concept clinical study 17 nerve transfers were performed in five patients. Nerve transfers included procedures to restore triceps function (N=4), wrist/finger/thumb extension (N=6) and finger flexion (N=7). Functional motor recovery (MRC ≥3) was achieved in 76% and 88% of transfers at 12 and 24 months, respectively.The preclinical study showed that FCS mimics epineurium and enable to repair nerve resembled to normal nerve tissue. Clinical study showed that patients received nerve transfer with FCS experienced consistent and early return of motor function in target muscles. These results provide proof of concept evidence that CND functions as an epineurial substitute and is promising for use in nerve transfer surgery.

Current Perspectives on Selective Dopamine D3 Receptor Antagonists/Partial Agonists as Pharmacotherapeutics for Opioid and Psychostimulant Use Disorders.

Over three decades of evidence indicate that dopamine (DA) D3 receptors (D3R) are involved in the control of drug-seeking behavior and may play an important role in the pathophysiology of substance use disorders (SUD). The expectation that a selective D3R antagonist/partial agonist would be efficacious for the treatment of SUD is based on the following key observations. First, D3R are distributed in strategic areas belonging to the mesolimbic DA system such as the ventral striatum, midbrain, and ventral pallidum, which have been associated with behaviors controlled by the presentation of drug-associated cues. Second, repeated exposure to drugs of abuse produces neuroadaptations in the D3R system. Third, the synthesis and characterization of highly potent and selective D3R antagonists/partial agonists have further strengthened the role of the D3R in SUD. Based on extensive preclinical and preliminary clinical evidence, the D3R shows promise as a target for the development of pharmacotherapies for SUD as reflected by their potential to (1) regulate the motivation to self-administer drugs and (2) disrupt the responsiveness to drug-associated stimuli that play a key role in reinstatement of drug-seeking behavior triggered by re-exposure to the drug itself, drug-associated environmental cues, or stress. The availability of PET ligands to assess clinically relevant receptor occupancy by selective D3R antagonists/partial agonists, the definition of reliable dosing, and the prospect of using human laboratory models may further guide the design of clinical proof of concept studies. Pivotal clinical trials for more rapid progression of this target toward regulatory approval are urgently required. Finally, the discovery that highly selective D3R antagonists, such as R-VK4-116 and R-VK4-40, do not adversely affect peripheral biometrics or cardiovascular effects alone or in the presence of oxycodone or cocaine suggests that this class of drugs has great potential in safely treating psychostimulant and/or opioid use disorders.

Masupirdine (SUVN‐502): A promising clinical candidate for the management of agitation in Alzheimer’s dementia

AbstractBackgroundMasupirdine is a potent and selective 5‐HT6 receptor antagonist and has completed Phase‐2 clinical proof of concept studies in patients with Alzheimer’s type dementia. 5‐HT6 receptors are G‐protein coupled receptors mainly localized in the brain regions primarily involved in cognition and behavior i.e., cortex, dorsal hippocampus and striatum. Evidence from the studies in animal models suggests that 5‐HT6 receptor antagonists may have a potential role in controlling neuropsychiatric behavior such as agitation.MethodMasupirdine has been evaluated in a phase‐2a proof‐of‐concept, 26‐week, randomized, double‐blind, placebo‐controlled, multicenter, parallel groups in United States under active US IND. Sub group analyses has been carried out to evaluate the effects of masupirdine on domains related to agitation/ aggression using Neuropsychiatric Inventory (NPI‐12) scale.Masupirdine was also tested in resident intruder task to evaluate its effects on animal model relevant to aggression. Additionally, masupirdine has been evaluated using microdialysis technique to assess its effect on the modulation of monoaminergic neurotransmitters.ResultIn Phase‐2 clinical study subgroup analyses, masupirdine showed robust efficacy in agitation/aggression scores in subjects who had baseline symptoms of agitation/aggression. The effect observed was greater than the minimum difference observed to be clinically important. Similar beneficial effects were observed on extended composite NPI‐12 score related to agitation/ aggression (agitation/ aggression + aberrant motor behavior + sleep and nighttime behavior disorder). Masupirdine was found to be safe and well tolerated in this study. In preclinical animal models, masupirdine reduced the aggressive behaviour in resident intruder task and increased neurotransmitters with role in modulation of agitation and aggression.ConclusionMasupirdine showed clinically meaningful improvement in the neuropsychiatric symptoms associated with agitation/aggression in patients with AD and reduced aggression in animal models. Masupirdine could be a differentiated novel therapeutic option for the management of agitation/ aggression in Alzheimer’s patients.

Topical application of Apremilast in the treatment of mild to moderate psoriasis

Mild to moderate psoriasis is highly prevalent in about 80% of the global psoriatic population. Current available treatment options for mild to moderate psoriasis are topical dosage forms and are associated variety of setbacks. To address these setbacks, Apremilast topical gels, 2% & 4%, w/w were developed, and a clinical proof of concept study (POC) was performed to establish efficacy and safety. A single centre randomized, double-blind, placebo-controlled study was conducted with apremilast topical gels 2% & 4% w/w in adult mild to moderate psoriatic patients for 12 weeks. The efficacy of the gels was evaluated by comparing the PASI scores before and after treatment of 12 weeks. Both gels exhibited a significant reduction in PASI values when compared with baseline PASI scores. An average percentage inhibition of PASI with test products, i.e. 2% and 4% w/w Apremilast topical gels, are about 46.8% and 34.6%, respectively, after 12 weeks of treatment. The results confirm that the apremilast topical gels are a good option for the treatment of mild to moderate psoriasis and have to be explored further.

Topical Application of Apremilast in the Treatment of Mild to Moderate Psoriasis

Background: Psoriasis is a chronic, autoimmune disorder that affects the skin and joints with an approximate global prevalence of 2–3%. Mild to moderate psoriasis is highly prevalent in about 80% of the global psoriatic population (2-3%). Currently available treatment options for mild to moderate psoriasis are topical dosage forms. Though a variety of topical formulations available, they are associated with different side effects. There is an unmet need for a product that can be used for a prolonged period with minimal side effects. Hence, Apremilast gel was developed and clinical proof of concept study (POC) was performed to investigate the efficacy and safety in mild to moderate psoriasis patients. Methods: A single-center randomized, double-blind, placebo-controlled study was conducted to evaluate the efficacy and safety of apremilast topical gels 2% & 4% w/w, in adult mild to moderate psoriatic patients for 12 weeks. Patients were examined at weeks 0, 2, 4, 8, and 12 weeks to assess the efficacy and safety. At 0 and 8 weeks, blood samples were collected to investigate the pharmacokinetics. The significance in % recovery was calculated statistically. Results: Both gels exhibited a significant reduction in PASI values when compared with baseline PASI scores. The average percentage inhibition of PASI with test products i.e. 2% and 4% w/w Apremilast topical gels is about 46.8% and 34.6% respectively after 12 weeks of treatment. Both the test products have not shown any adverse effects, hematological & biochemical changes and have exhibited Cmax less than 20ng/ml after 6 hours of application. Conclusion: Results have shown that topically applied apremilast could be an effective and safe option for the management of mild to moderate psoriasis.

Abstract 4276: Target enrichment to enable RNA-sequencing on FFPE samples: Application to a patient selection biomarkers in colorectal cancer

Abstract In proof of concept clinical studies, selecting for patients that are the most likely to respond to a given therapy will increase the chance of demonstrating clinical benefit for a new investigational agent. Archived Formalin-Fixed, Paraffin-Embedded (FFPE) tissue taken at the time of first biopsy and cancer diagnosis are a ready source of material for such patient selection assays; however if the selection assay is RNA-based sequencing based assay, FFPE tumor tissues are typically refractory to standard methods. Here we describe Target Enrichment, an RNA-sequencing technique that enabled analysis of FFPE tumor tissue. The Target Enrichment technique provides the necessary, high quality gene expression data required for identifying the mesenchymal subtype of CRC. In this experiment, RNA was extracted from a set of 192 histologically characterized, archived colorectal cancer (CRC) FFPE tumor tissues with sufficient representation of all Consensus Molecular Subtypes of CRC. A major issue with FFPE tissue is that nucleic acids are not well preserved in the fixation process. The RNA extraction method was carefully optimized in advance toward improving yield and integrity of the total RNA. In order to accommodate the low FFPE RNA integrity, we used Target Enrichment, a technique derived from whole-exome sequencing that isolates and amplifies library fragments corresponding to only the transcribed regions of interest. After testing several protocols, we established a Target Enrichment technique to remove low-quality library fragments that would otherwise interfere and impair sequencing quality. This method allowed us to generate high-quality sequencing data for samples that proved challenging using traditional RNA-seq methods. Fresh-frozen tissue samples were used as a benchmark and our Target Enrichment technique showed comparable performance across multiple sequencing quality metrics including mapping rate, expression profiling efficiency, number of genes detected, also the pass rate. Furthermore, data generated using our Target Enrichment technique aligned with the known frequencies of the Consensus Molecular Subtypes of CRC. Overall, our Target Enrichment technique provides a protocol for generating high quality RNA-seq libraries from highly degraded FFPE RNA. Citation Format: Maximilian Rogers-Grazado, Joachim Theilhaber, Jack Pollard, Alexei Protopopov. Target enrichment to enable RNA-sequencing on FFPE samples: Application to a patient selection biomarkers in colorectal cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4276.

The anti-ageing effects of a natural peptide discovered by artificial intelligence.

As skin ages, impaired extracellular matrix (ECM) protein synthesis and increased action of degradative enzymes manifest as atrophy, wrinkling and laxity. There is mounting evidence for the functional role of exogenous peptides across many areas, including in offsetting the effects of cutaneous ageing. Here, using an artificial intelligence (AI) approach, we identified peptide RTE62G (pep_RTE62G), a naturally occurring, unmodified peptide with ECM stimulatory properties. The AI-predicted anti-ageing properties of pep_RTE62G were then validated through in vitro, ex vivo and proof of concept clinical testing. A deep learning approach was applied to unlock pep_RTE62G from a plant source, Pisum sativum (pea). Cell culture assays of human dermal fibroblasts (HDFs) and keratinocytes (HaCaTs) were subsequently used to evaluate the in vitro effect of pep_RTE62G. Distinct activities such as cell proliferation and ECM protein production properties were determined by ELISA assays. Cell migration was assessed using a wound healing assay, while ECM protein synthesis and gene expression were analysed, respectively, by immunofluorescence microscopy and PCR. Immunohistochemistry of human skin explants was employed to further investigate the induction of ECM proteins by pep_RTE62G ex vivo. Finally, the clinical effect of pep_RTE626 was evaluated in a proof of concept 28-day pilot study. In vitro testing confirmed that pep_RTE62G is an effective multi-functional anti-ageing ingredient. In HaCaTs, pep_RTE62G treatment significantly increases both cellular proliferation and migration. Similarly, in HDFs, pep_RTE62G consistently induced the neosynthesis of ECM protein elastin and collagen, effects that are upheld in human skin explants. Lastly, in our proof of concept clinical study, application of pep_RTE626 over 28days demonstrated anti-wrinkle and collagen stimulatory potential. pep_RTE62G represents a natural, unmodified peptide with AI-predicted and experimentally validated anti-ageing properties. Our results affirm the utility of AI in the discovery of novel, functional topical ingredients.

Lactobacillus reuteri DSM 17938 as a Novel Topical Cosmetic Ingredient: A Proof of Concept Clinical Study in Adults with Atopic Dermatitis.

Atopic Dermatitis (AD) is a chronically relapsing skin condition characterized by dry, itchy, and inflamed skin where sufferers can frequently be subject to infections. Probiotics are known to be potent immune-modulators, and live Lactobacillus reuteri DSM 17938 has shown to be anti-inflammatory but also to possess antimicrobial and barrier function properties. This study aimed to investigate and compare two investigational ointment products (topical probiotic and control) for cutaneous acceptability, safety, and efficacy under normal conditions of use, in adult subjects with atopic dermatitis. The products were applied twice daily for 8 weeks, and cutaneous acceptability, SCORAD index, local SCORAD, and adverse events were evaluated after 4 and 8 weeks of treatment. At the end of the observations, it was demonstrated that both the probiotic-containing and probiotic-free ointments were both cutaneously acceptable and safe. It importantly showed a statistically and clinically significant improvement of the SCORAD index and local SCORAD in adult subjects with AD after 4 and 8 weeks of continuous use. In conclusion, we show evidence that the probiotic product, containing live L. reuteri DSM 17938 as an extra ingredient, is safe and promising as a novel topical cosmetic ointment and with further testing could be a standard topical product for the management of atopic dermatitis or other disorders associated with the skin.

Digital Therapeutic Platform for Management of Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a complex, multi-system autoimmune disease of unclear etiology that causes significant morbidity and, in severely affected patients, early mortality. Despite efforts from academic and private research entities, pharmaceutical companies, and patient advocacy groups, and hundreds of millions of dollars in spending, numerous gaps in care still exist. A digital therapeutic platform is described that uses self-tracking technology, analytics, and telehealth coaching to identify and remove possible dietary and/or other lifestyle triggers of SLE. A clinical proof of concept study was performed with 18 SLE patients over a 12 week program. All participants reported improvements in their symptoms, including pain, fatigue, digestive, and other physical symptoms.Clinical Relevance- This study demonstrates the technical and clinical feasibility of a digital therapeutic platform to improve the health-related quality of life in patients with systemic lupus erythematosus.

The dilemma of renin-angiotensin system inhibitors in coronavirus disease 2019 (COVID-19): insights into lung fluid handling and gas exchange in heart failure patients.

The dilemma of renin-angiotensin system inhibitors in coronavirus disease 2019 (COVID-19): insights into lung fluid handling and gas exchange in heart failure patients.

459P - Proof of concept clinical study by US-guided intratumor injection of VCN-01, an oncolytic adenovirus expressing hyaluronidase in patients with pancreatic cancer

459P - Proof of concept clinical study by US-guided intratumor injection of VCN-01, an oncolytic adenovirus expressing hyaluronidase in patients with pancreatic cancer

Advances in Antiwolbachial Drug Discovery for Treatment of Parasitic Filarial Worm Infections

The intracellular bacteria now known as Wolbachia were first described in filarial worms in the 1970s, but the idea of Wolbachia being used as a macrofilaricidal target did not gain wide attention until the early 2000s, with research in filariae suggesting the requirement of worms for the endosymbiont. This new-found interest prompted the eventual organization of the Anti-Wolbachia Consortium (A-WOL) at the Liverpool School of Tropical Medicine, who, among others have been active in the field of antiwolbachial drug discovery to treat filarial infections. Clinical proof of concept studies using doxycycline demonstrated the utility of the antiwolbachial therapy, but efficacious treatments were of long duration and not safe for all infected. With the advance of robotics, automation, and high-speed computing, the search for superior antiwolbachials shifted away from smaller studies with a select number of antibiotics to high-throughput screening approaches, centered largely around cell-based phenotypic screens due to the rather limited knowledge about, and tools available to manipulate, this bacterium. A concomitant effort was put towards developing validation approaches and in vivo models supporting drug discovery efforts. In this review, we summarize the strategies behind and outcomes of recent large phenotypic screens published within the last 5 years, hit compound validation approaches and promising candidates with profiles superior to doxycycline, including ones positioned to advance into clinical trials for treatment of filarial worm infections.

Next tier in vitro and in vivo nonclinical studies further elucidating the safety and toxicity profile of MB-102, a novel fluorescent tracer agent for measurement of glomerular filtration rate

MB-102 was designed for measurement of real-time glomerular filtration rate (GFR). Previously reported in vitro and in vivo nonclinical studies clearly demonstrated negligible toxicity, resulting in FDA clearance for First-in Human, proof of concept clinical studies. The next tier of safety and toxicity studies are reported herein.MB-102 did not demonstrate any phototoxic potential in a BALB/c 3T3 mouse fibroblast study. Co-administration of MB-102 and iohexol resulted in pharmacokinetic parameters virtually identical to the values observed upon individual administration in beagle dogs. A single dose of MB-102 administered either intravenously (18.6 mg/mL) or perivenously (0.25 mL) was well-tolerated in NZ white rabbits, with no adverse inflammation or irritation. MB-102 did not induce micronuclei in polychromatic erythrocytes for rat bone marrow cells treated up to 450 mg/kg/day, the maximum feasible dose. Two separate optical imaging studies demonstrated that MB-102 distributes rapidly and thoroughly throughout the test subjects, followed by rapid clearance from the body without any preferential localization in any particular tissue or organ, with the exception of the bladder, which is totally consistent with a known GFR agent. In addition, two-week repeat intravenous (once-daily) toxicity and toxicokinetic studies were conducted in rats and beagles, with no MB-102- related effects.Thus, for the studies reported herein, there were no toxicological effects of concern for MB-102.

622 A proof of concept clinical study to test the efficacy and safety of VB1953 in moderate to severe acne subjects

622 A proof of concept clinical study to test the efficacy and safety of VB1953 in moderate to severe acne subjects

The importance of inhibition of a catabolic pathway of methotrexate metabolism in its efficacy for rheumatoid arthritis

Methotrexate (MTX), an antifolate, is the anchor drug for the treatment of rheumatoid arthritis (RA). It is inexpensive, effective, and generally safe. When clinical response is inadequate, biological therapies are commonly used in combination with MTX. However, biological agents have safety concerns (i.e. infections, malignancy) and the addition of a biologic agent is expensive, making strategies to improve MTX efficacy important.Inhibition of pathways of folate metabolism involving purine metabolism by MTX, have been traditionally emphasized as important in MTX efficacy. However, inhibition MTX catabolism may also be important. MTX is irreversibly hydroxylated to form 7-hydroxy methotrexate (7-OH-MTX) by aldehyde oxidase (EC 1.2.3.1) (AOX). Catabolism of MTX to 7-OH-MTX is the first metabolic process imposed on an oral dose of MTX and will alter subsequent interactions of MTX with other enzymes. 7-OH-MTX is less potent than MTX in the treatment of rat adjuvant arthritis. RA patients with a low capacity to catabolize MTX to 7-OH-MTX do better clinically than individuals who are rapid formers of 7-OH-MTX. Therefore, altering the catabolism of MTX may be an innovative way to improve MTX efficacy.Raloxifene is a FDA-approved therapy for postmenopausal osteoporosis and for the reduction of invasive breast cancers but has no known activity in RA. Raloxifene is a potent inhibitor of human liver AOX. Postmenopausal women with RA frequently have low bone mineral density and would be candidates for raloxifene and MTX combination therapy. The effect of raloxifene on MTX metabolism has never been studied.Our hypothesis is that in postmenopausal women with RA and osteoporosis treated with MTX and raloxifene, the inhibition of AOX with resultant decreased formation of 7-OH MTX; will increase MTX levels and improve MTX efficacy. This hypothesis could be studied in an open-label, proof of concept clinical study in individuals before and after the addition of raloxifene. Red blood cell MTX and 7-OH-MTX levels and RA disease activity (DAS28) would be measured. In possible future studies, there are dietary substances, as supplements, (e.g. epigallocatechin gallate in green tea and resveratrol) which inhibit human liver AOX which could be evaluated.

Naftazone in advanced Parkinson's disease: An acute L-DOPA challenge randomized controlled trial

IntroductionThere is an unmet need to better control motor complications in Parkinson's disease (PD). Naftazone, which exhibits glutamate release inhibition properties, has shown antiparkinsonian and antidyskinetic activity in preclinical models of PD and in a clinical proof of concept study. MethodsWe conducted a double-blind randomized placebo-controlled cross-over trial in PD patients with motor fluctuations and dyskinesia testing naftazone 160 mg/day versus placebo for 14 days. The two co-primary endpoints were the area under curve (AUC) of motor (MDS-UPDRS part III) and dyskinesia (AIMS) scores during an acute levodopa challenge performed at the end of each period. Secondary endpoints were UDysRS and axial symptoms scores during the challenge; AIMS, UDysRS, and time spent with or without dyskinesia the day before the challenge. The primary analysis was performed in the per protocol population. ResultsSixteen patients were included in the analysis. There was no difference between naftazone and placebo for the AUC of MDS-UPDRS III (−89, 95%CI[-1071; 893], p = 0.85), and AIMS (70, 95%CI[-192; 332], p = 0.57). At the end of treatment periods, AIMS score tended to be lower with naftazone than placebo (4.4 ± 3.4 versus 6.7 ± 4.4, p = 0.07), but UDysRS scores and other secondary outcomes were not different. Naftazone was safe and well tolerated. ConclusionsThis study did not confirm previous results on the efficacy of naftazone on dyskinesia nor motor fluctuations highlighting the problem of translating results obtained in preclinical models into clinical trials. Further investigation of naftazone may be conducted in PD with longer treatment duration.

Abstract CT137: Biomarkers of response to leukemia stem cell targeted therapy with low dose daunorubicin in relapsed/refractory acute leukemia

Abstract Patients with acute leukemia harbor chemotherapy-resistant leukemia stem cells (LSC) which ultimately lead to disease relapse in many patients. Clinical trial design of LSC targeted therapy is a challenge. The majority of leukemia cells are differentiated leukemia cells, with LSC comprising <1%. Clinical benefit of LSC targeted therapy is best measured by overall survival and difficult to measure in the short-term using the conventional endpoint of remission. Response to LSC targeted therapy may be confounded by lack of response of the bulk population. Optimal treatment strategies may ultimately include combined or sequential therapy targeted at both the LSC and bulk leukemia cell populations. However, this approach makes it difficult to measure effects on LSC specifically. In order to isolate specific effects of targeted therapy on LSC, we designed a clinical proof of concept study using serial bone marrow aspirates to measure the effects of LSC-targeted therapy in adults with relapsed acute leukemia. In pre-clinical models, we demonstrated that AKT-β-catenin interaction, indicated by Akt phosphorylation of β-catenin at serine 552 (pS552) is critical for LSC self-renewal. High throughput screening identified daunorubicin (DNR) as a potent inhibitor of pS552. To better understand the role of DNR in specifically targeting LSC, we experimented with lower doses, recognizing that most leukemia patients receive high dose DNR, yet LSC persist and result in disease relapse. In our murine model, low dose anthracycline inhibited LSC number and function. We hypothesized that low dose DNR treatment inhibits LSC number in patients with acute leukemia. Our pilot trial includes relapsed acute myeloid (AML) and lymphoblastic leukemia (ALL) patients (NCT02914977). Bone marrow aspiration was performed at study entry. Patients received DNR at 6.75 mg/m2/day x 5 days followed by repeat bone marrow aspiration on day 8. The plasma pharmacokinetics of low dose DNR was also characterized. Samples from pre- and post-treatment were analyzed by flow cytometry for the frequency (%) of total cells that are pS552-β-catenin positive LSCs (defined as CD34+ CD38- cells that express TIM3, which distinguishes LSCs from normal stem cells). Nine patients (8 AML, 1 ALL) have been treated. Seven patients had previous DNR treatment. Two patients had prior allogeneic stem cell transplant. Five of 9 patients had reduction in LSC number as measured by flow cytometry; 3/5 had prior DNR treatment. In this pilot proof of concept trial, we demonstrate the feasibility of serial bone marrow aspiration to measure biomarkers of LSC response. In our small series, 5/9 relapsed leukemia patients had reduction in LSC number after one cycle of low dose DNR. Future plans include incorporating low dose DNR following conventional chemotherapy in newly diagnosed patients with AML. This clinical proof of concept trial design with serial bone marrow aspirations may also be used to rapidly assess the validity of other LSC markers and targeted agents. Citation Format: Tara L. Lin, John M. Perry, Xi He, Gregory Reed, Na Zhang, Scott Weir, Joseph P. McGuirk, Linheng Li. Biomarkers of response to leukemia stem cell targeted therapy with low dose daunorubicin in relapsed/refractory acute leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT137.