Production and Quality Control of [177Lu]Lu-PSMA-I&T: Development of an Investigational Medicinal Product Dossier for Clinical Trials

Since prostate cancer is the most commonly diagnosed malignancy in men, the theranostic approach has become very attractive since the discovery of urea-based PSMA inhibitors. Different molecules have been synthesized starting from the Glu-urea-Lys scaffold as the pharmacophore and then optimizing the linker and the chelate to improve functional characteristics. This article aimed to highlight the quality aspects, which could have an impact on clinical practice, describing the development of an Investigational Medicinal Product Dossier (IMPD) for clinical trials with [¹⁷⁷Lu]Lu-PSMA-I&T in prostate cancer and other solid tumors expressing PSMA. The results highlighted some important quality issues of the final preparation: radiolabeling of PSMA-I&T with lutetium-177 needs a considerably longer time compared with the radiolabeling of the well-known [¹⁷⁷Lu]Lu-PSMA-617. When the final product was formulated in saline, the stability of [¹⁷⁷Lu]Lu-PSMA-I&T was reduced by radiolysis, showing a decrease in radiochemical purity (<95% in 24 h). Different formulations of the final product with increasing concentrations of ascorbic acid have been tested to counteract radiolysis and extend stability. A solution of 20 mg/mL of ascorbic acid in saline prevents radiolysis and ensures stability over 30 h.     

The 160.44 day Lu as a new gamma calibration standard

Gamma emissions in the decay of 160.44 day isomer ^177mLu were analyzed with a 60 cc, coaxial HPGe detector. The energies and intensities of 50 gamma transition - 40 of which belong to the daughter ¹⁷⁷Hf, that was excited by beta decay of the isomer, and 10 to the isomeric transition in ¹⁷⁷Lu were precisely determined. These precise values have resulted in an internally consistent decay scheme for ^177mLu. These gamma intensity and gamma emission probability measurements will help in making the long lived ^177mLu available as a calibration standard for HPGe detectors and also for other applications.     

On the practical aspects of large-scale production of 177Lu for peptide receptor radionuclide therapy using direct neutron activation of 176Lu in a medium flux research reactor: the Indian experience

This paper accentuates on the practical aspects and intricate technicalities involved in the large-scale production of ¹⁷⁷Lu with specific activity >740 GBq mg^?1 following (n,γ)¹⁷⁷Lu route in a medium flux (~1.2 × 10¹⁴ n cm^?2 s^?1 thermal neutron) research reactor. The implication of target burn-up on the specific activity of ¹⁷⁷Lu during irradiation was discussed in detail. ¹⁷⁷Lu obtained from this route has been extensively utilized for targeted therapy in patients with neuroendocrine tumor in India. The important details available from our experience, as well as technical know-how, would be of considerable value for institutions planning to pursue ¹⁷⁷Lu production through (n,γ)¹⁷⁷Lu route.     

Single vial formulation for theranostic radiopharmaceutical preparation

The present work was aimed at development of pharmaceutical grade single vial kit like formulation of somatostatin analogue, DOTA–Tyr³–Thr⁸-Octreotide (DOTATATE) suitable for radiolabeling with both diagnostic (⁶⁸Ga) and therapeutic (¹⁷⁷Lu) radioisotope. Single vial kit like formulation of DOTATATE was prepared. Radiolabeling methods with ⁶⁸Ga and ¹⁷⁷Lu were standardized. The pharmaceutical purity and stability of formulation was studied over a period of 6 months. Pharmacokinetics of radiolabeled preparations was studied in Swiss mice. DOTATATE formulation with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer was successfully prepared. Both ⁶⁸Ga–DOTATATE and ¹⁷⁷Lu–DOTATATE complexes were formed with >95 % radiochemical purity. Biodistribution studies of ⁶⁸Ga–DOTATATE and ¹⁷⁷Lu–DOTATATE complexes in Swiss mice revealed fast clearance of activity via renal route. Single vial kit like formulation suitable for easy preparation of ⁶⁸Ga–DOTATATE and ¹⁷⁷Lu–DOTATATE at hospital radiopharmacy was successfully demonstrated.     

Radiochemical Feasibility of Mixing of 99mTc-MAA and 90Y-Microspheres with Omnipaque Contrast

Yttrium-90 (⁹⁰Y) microspheres are widely used for the treatment of liver-dominant malignant tumors. They are infused via catheter into the hepatic artery branches supplying the tumor under fluoroscopic guidance based on pre-therapy angiography and Technetium-99m macroaggregated albumin (^99mTc-MAA) planning. However, at present, these microspheres are suspended in radiolucent media such as dextrose 5% (D5) solution. In order to monitor the real-time implantation of the microspheres into the tumor, the ⁹⁰Y microspheres could be suspended in omnipaque contrast for allowing visualization of the correct distribution of the microspheres into the tumor. The radiochemical purity of mixing ⁹⁰Y-microspheres in various concentrations of omnipaque was investigated. The radiochemical purity and feasibility of mixing ^99mTc-MAA with various concentrations of a standard contrast agent were also investigated. Results showed the radiochemical feasibility of mixing ⁹⁰Y-microspheres with omnipaque is radiochemically acceptable for allowing real-time visualization of radioembolization under fluoroscopy.     

Investigation of bacterial adherence to a non-precious alloy with radiolabeling method

Summary The objective of this study was to investigate the bacterial adherence to a non-precious alloy with radiolabeling method. S. mutans, E. coliand C. albicanswere labeled with ^99mTc by using stannous chloride and their radiolabeling yields were calculated. After the labeling procedure, metal disks (3 mm×10 mm) were treated with microorganisms. The amount of labeled microorganisms adhered on metal surfaces was determined by activity measurements. The labeling yields for S. mutans, E. coliand C. albicanswere 69.95±7.58%, 78.84±0.44% and 79.71±10.17%, respectively. The mean values for adherence for S. mutans, E. coliand C. albicanson metal samples were 7.02±2.18%, 0.96±0.49% and 8.80±8.24%, respectively. The radiolabeling method could be considered as safe and precise for determining the adherence of microorganisms.     

Accurate determination of half-life and radionuclidic purity of reactor produced <Superscript>177g</Superscript>Lu (<Superscript>177m</Superscript>Lu) for metabolic radiotherapy

Thanks to its favorable decay characteristics, ^177gLu is finding several applications in nuclear medicine, especially for palliative metabolic radiotherapy of cancer and radioimunotherapy. ^177gLu is produced in thermal nuclear reactor either by direct neutron capture ¹⁷⁶Lu(n,γ)^177(m+g)Lu on either natural or enriched ¹⁷⁶Lu target, or by reaction on enriched ¹⁷⁶Yb target followed by negatron decay. The latter method does produce a high radionuclidic purity and high specific activity radionuclide in no-carrier-added form, since ¹⁷⁷Yb decays solely to the ground state ^177gLu. Conversely, the first method does produce a low specific activity ^177gLu in carrier-added form,¹ contaminated by the long-lived radioisotopic impurity ^177mLu. The accurate determination of radionuclidic purity and half-life of ^177gLu carried out by HPGe and LSCS is presented in some details.     

Radionuclide purity assessment and calibration of <Superscript>90</Superscript>Y(<Superscript>177</Superscript>Lu) glass particles using gamma-ray spectrometry

Intra-hepatic administration of radioactive glass microspheres is a treatment for patients with primary liver cancer and hepatic metastases. The purpose of this study was radionuclide purity assessment of new glass particles containing two radionuclide, ⁹⁰Y as a therapeutic source and also ¹⁷⁷Lu as a source of diagnostic gamma. For the mixed source, activity measurement using a dose calibrator cannot be used and we need new calibration methods. YAS (Yb) and YAS compositions were sol–gel derived glass particles and production of ⁹⁰Y (¹⁷⁷Lu) and ⁹⁰Y particles was performed using the Tehran Research Reactor. The radionuclide purity was carried out using γ-spectrometry with HPGe detector. A non-destructive spectroscopic assay was employed due to a newly updated low uncertainty positron branching ratio of ⁹⁰Y that emit 511 keV annihilation radiations. In another method, a new calibration of ⁹⁰Y using a non-destructive spectroscopic assay of ⁸⁸Y were investigated. Potential radionuclide impurity include: ⁸⁸Y, ¹⁵²Eu, ⁶⁰Co with activity 100, 50 and 5 Bq per 1 mg of that are not harmful for patients due to delivering radioactive particles about 20–50 mg in ⁹⁰Y(¹⁷⁷Lu) glass microspheres. Among of radionuclide impurity, ¹⁵²Er with a half life of 13.54 years and ⁸⁸Y with a half life of 106.65 days was important in the residual delivery device. For calibration of ⁹⁰Y with monitoring of 511 keV, errors were12.2–21%. In calibration of ⁹⁰Y using gamma spectroscopic assay of ⁸⁸Y, there was an error less than 14%. Spectroscopic assay of ⁸⁸Y can be performed easily and has more repeat for our purpose.     

Production of no-carrier-added 177Lu via the 176Yb(n, &;#947;)177Yb &;#8594; 177Lu process

No-carrier-added ¹⁷⁷Lu was produced by the ¹⁷⁶Yb(n,)¹⁷⁷Yb¹⁷⁷Lu process using enriched ¹⁷⁶Yb₂O₃. The radiochemical separation of the nca ¹⁷⁷Lu from the macroscopic ytterbium target was investigated by reversed-phase ion-pair HPLC. Effects of the concentrations of 2-hydroxyisobutyric acid and 1-octanesulfonate in the eluent, the amount of Yb₂O₃, the type and length of the C₁₈ column on the separation efficiency were examined. Under optimum conditions, the nca ¹⁷⁷Lu was obtained in radiochemically pure form from 5 mg of Yb₂O₃ with a separation yield of 84%.     

Separation of carrier-free lutetium-177 from neutron irradiated natural ytterbium target

Neutron irradiation of naturally occurring Yb produces small amounts of carrier-free¹⁷⁷Lu activity. Cation exchange chromatography in the displacement development mode using Dowex-50X8, 200–400 mesh resin, and Zn²⁺ as the separating ion was used to separate¹⁷⁷Lu produced in a neutron irradiated Yb target. 0.04M -hydroxyisobutyric acid at pH 4.6±2 and temperature 26±1°C was used to elute carrier-free¹⁷⁷Lu in 70% yield and at a radionuclidic purity greater than 99%.     

Reactor produced 177Lu of specific activity and purity suitable for medical applications

Four production methods of ¹⁷⁷Lu, e.g. from natural and enriched Lu and from natural and enriched Yb were considered and experimentally evaluated. The samples of all 4 materials were irradiated in a nuclear reactor, the activity of ¹⁷⁷Lu measured and compared with a computed one. In the case of ¹⁷⁷Lu produced from enriched Lu target the amount of activity obtained is 40 to 70% higher than calculated. The results achieved will be applied for the optimization of ¹⁷⁷Lu production for medical applications.     

Recovery of 177Lu from Irradiated HfO2 Targets for Nuclear Medicine Purposes

A new method of production of one of the most widely used isotopes in nuclear medicine, ¹⁷⁷Lu, with high chemical purity was developed; this method includes irradiation of the HfO₂ target with bremsstrahlung photons. The irradiated target was dissolved in HF and then diluted and placed onto a column filled with LN resin. Quantitative sorption of ¹⁷⁷Lu could be observed during this process. The column later was rinsed with the mixture of 0.1 M HF and 1 M HNO₃ and then 2 M HNO₃ to remove impurities. Quantitative desorption of ¹⁷⁷Lu was achieved by using 6 M HNO₃. The developed method of ¹⁷⁷Lu production ensures high purification of this isotope from macroquantities of hafnium and zirconium and radioactive impurities of carrier-free yttrium. The content of ^177mLu in ¹⁷⁷Lu in photonuclear production was determined. Due to high chemical and radionuclide purity, ¹⁷⁷Lu obtained by the developed method can be used in nuclear medicine.     

Preparation,quality control and stability of <Superscript>99m</Superscript>Tc-cefuroxime axetil

Cefuroxime axetil, a cephalosporin antibiotic used to treat bacterial infections, was investigated to label with ^99mTc. Radiolabeling of cefuroxime axetil was carried out by using stannous chloride method. Effects of pH and stannous chloride amount on the radiolabeling yield were investigated. The radiochemical purity of ^99mTc-cefuroxime axetil was determined by thin layer radio chromatography (TLRC), electrophoresis and high performance liquid chromatography. The maximum radiolabeling yield was 98±1%.     

Low-temperature heat capacity and high-temperature thermal behavior of sodium lutetium molybdate phosphate Na<Subscript>2</Subscript>Lu(MoO<Subscript>4</Subscript>)(PO<Subscript>4</Subscript>)

The low-temperature heat capacity of Na₂Lu (MoO₄)(PO₄) was measured by adiabatic calorimetry in the range of 7.47–345.74 K. The experimental data were used to calculate the thermodynamic functions of Na₂Lu (MoO₄)(PO₄). At 298.15 K, the following values were obtained: C _p ⁰ (298.15 K) = 237.7 ± 0.1 J/(K mol), S ⁰(298.15 K) = 278.1 ± 0.8 J/(K mol), H ⁰(298.15 K) ? H ⁰ (0 K) = 42330 ± 20 J/mol, and Φ⁰(298.15 K) = 136.1 ± 0. 3 J/(K mol). A heat capacity anomaly was found in the range of 10-67 K with a maximum at T _tr = 39.18 K. The entropy and enthalpy of transition are ΔS = 12.39 ± 0.75 J/(K mol) and ΔH = 403 ± 16 J/mol. The thermal investigation of sodium lutetium molybdate phosphate in the high-temperature range (623–1223 K) was performed using differential scanning calorimetry. It was found that during melting in the range of 1030–1200 K, Na₂Lu(MoO₄)(PO₄) degrades to simpler compounds; the degradation scenario is verified by X-ray powder diffraction.     

186Re/188Re labeled polypeptide microspheres as a potential radiation synovectomy agent

Polypeptide microspheres containing polycysteine crosslinked with polylysine were prepared and radiolabeled with¹⁸⁶Re and¹⁸⁸Re. High labeling yields with the microspheres with both¹⁸⁶Re and¹⁸⁸Re (97%) were obtained, and above 99% retention of radiolabels in water in 24 hours was obtained. Rhenum-186 labeled polycysteine and polylysine microspheres (11 ratio, 20 m as mean diameter) were injected intra-articularly into the rear stifes (knee joints) of normal New Zealand white rabbits. About 87% of injected dose was retained in rabbit stifles and adjacent tissues in 96 hours after injection, while most of the activity lost from the joints was excreted in the urine. Due to its simplicity of preparation and radiolabeling, versatility, and biodegradability, this type of conjugate system may become the therapeutics of choice for radiation synovectomy.     

Preparation and quality control and biodistribution studies of [90Y]-DOTA-cetuximab for radioimmunotherapy

Yttrium-90 is a useful radionuclide for radioimmunotherapy (RIT) and the anti-epidermal growth factor receptor (anti-EGFR) antibody cetuximab is clinicsally approved for the treatment of EGFR-expressing metastatic colorectal cancer and advanced head and neck cancer. Thus in this work radiolabeling of monoclonal anti-EGFR with ⁹⁰Y for radioimmunotherapy (RIT) is targetted. Cetuximab was successively labeled with [⁹⁰Y] chloride (74 MBq) 2 mCi after conjugation with macrocyclics bifunctional chelating agent, 1,4,7,10-tetraazacyclododecane-N,N′,N″,N″′-tetraacetic acid mono-(N-hydroxysuccinimidyl) ester (DOTA-NHS), purified and concentrated by centrifugation using an Amicon Ultra-15 filter (Millipore, MWCo, 30000). ⁹⁰Y chloride was obtained by ⁹⁰Sr/⁹⁰Y generator. Radiolabeling was completed in 2 h by the addition of DOTA-cetuximab conjugate at 42 °C. The stability of radiolabeled was studied in human serum. Biodistribution studies in normal rats were carried out to determine the radioimmunoconjugate distribution up to 96 h. Radiochemical purity of 92 % (using ITLC) was obtained for final radioimmunoconjugate (Specific activity = 0.55 GBq/mg). Stability of radiolabeled protein in presence of human serum was tested at 37 °C for up to 24 h. Biodistribution studies demonstrated the highest ID/g % in the blood (2.62 ± 0.005 at 24 h) and the liver (2.19 ± 0.001). This study demonstrated that ⁹⁰Y-DOTA-cetuximab is a potential compound for the treatment of EGFR-expressing cancers.     

Screening for a 177Lu-labeled CA19–9 monoclonal antibody via PET imaging for colorectal cancer therapy

Carbohydrate antigen 19–9 (CA19–9) with multi epitopes relatively high expresses on colorectal cancer (CRC) cells, making it an attractive target for developing radioimmunotherapy (RIT) for CRC. The lutetium-177 (¹⁷⁷Lu) labeled monoclonal antibodies (mAbs) can selectively bind the corresponding antigens and release targeted cytotoxic radiation, which could induce cell apoptosis and reduce the drug-induced resistance. Here, a series of CA19–9 mAbs were labeled with zirconium-89 (⁸⁹Zr), and one with high tumor uptake was screened via PET imaging, which has potential application for the diagnosis of CRC. Then the screened mAb (C003) labeled with ¹⁷⁷Lu was utilized for CA19–9 targeted RIT, which presents a significant suppression effect on the growth of colo205 xenografts than immunotherapy alone. Meanwhile, the side effects of ¹⁷⁷Lu-DOTA-C003 are limited according to the results of in vivo study. Both ⁸⁹Zr-DFO-C003 for CRC immune-PET imaging and ¹⁷⁷Lu-DOTA-C003 for RIT against CRC exhibit good potential in clinical applications.     

Aspects of yield and specific activity of (n,γ) produced 177Lu used in targeted radionuclide therapy

¹⁷⁷Lu-labeled receptor avid peptides and monoclonal antibodies have been effectively used in targeted tumor therapy, owing to the ideally suited decay properties and favourable production logistics of ¹⁷⁷Lu [T_½ = 6.65 days; E_β(max) = 497 keV (78.6 %); E_γ = 208 keV (11.0 %)]. The specific activity of ¹⁷⁷Lu produced by the (n,γ) route is one of the important criteria, which determines the efficacy of ¹⁷⁷Lu-labeled receptor-avid biomolecules. The present article highlights that the specific activity of (n,γ) produced ¹⁷⁷Lu cannot be calculated by simply dividing the produced activity by the mass of the target irradiated, unlike other (n,γ) produced medical radioisotopes and there is a significant enhancement of specific activity due to the burn up of the Lu target during irradiation, which is an added advantage towards the utilization of ¹⁷⁷Lu in receptor specific therapeutic radiopharmaceuticals.     

Formulation and evaluation of quince seeds mucilage – sodium alginate microspheres for sustained delivery of cefixime and its toxicological studies

Quince seed mucilage was used in a combination of sodium alginate to develop sustained-release microspheres of cefixime. Physical characterizations such as FTIR, TGA, DSC, and SEM were performed on the prepared microspheres. The swelling of microspheres was maximum at pH 7.4 and reduced at acidic pH. The average particle size ranged from 679 µm ± 0.21 to 810 µm ± 0.31, while the drug encapsulation efficiency range was found as 73.76 ± 0.24–85.63 ± 0.46. In vitro release profile of QSM-alginate-cefixime microspheres followed Korsmeyer-Peppas model (R² = 0.9732-–0.9946); and release was non-Fickian as we found value of n > 1. This study reveals the benefits of QSM-alginate microspheres for the sustained release of cefixime without any toxicity and it also improved antibacterial properties.     

Radioiodination and biodistribution of isolated lawsone compound from Lawsonia inermis (henna) leaves extract

Lawsonia inermis (henna) is one of the most effective medicinal plants and it has been using for treatment of wounds and burns for centuries. The using of Henna leaves is very popular for cosmetic as well as medicine in many countries. Henna leaves contain lots of different compounds and lawsone (LW) is the main one. In current study, extraction with bidistillated water of henna leaves was performed and LW was isolated by using high performance liquid chromatography system. Chemical structure of LW was evaluated by nuclear magnetic resonance method. LW was radiolabeled with iodine-131 (¹³¹I) radionuclide which is well known for nuclear imaging and therapy in nuclear medicine by utilizing iodogen method. The yield of radiolabeling of LW (¹³¹I-LW) was calculated as 92.70 ± 4.312 % (n = 10) by thin layer radio chromatography. Its in vivo biological activity was investigated by biodistribution studies which were performed by using healthy female and male Balb/C mice. According to results of biodistribution, uptake of ¹³¹I labeled LW compound in uterus, breast and ovary for female mice and prostate in male mice was higher than other organs in the body.