The Molecule

A radioligand has three modules. The ligand binds an antigen on the surface of the tumor cell. A chelator (a ring of donor atoms which holds a metal ion) carries the radiometal, and a linker joins the two and sets the charge and clearance of the whole molecule. Since the chelator binds both gallium and lutetium, the same molecule is used for imaging when it is labeled with ⁶⁸Ga and for therapy when it is labeled with ¹⁷⁷Lu.

TUMOR CELL M ⁶⁸Ga Imaging PET scan confirms the target before therapy ¹⁷⁷Lu Therapy β⁻ emitter, 6.65 d, imageable by SPECT α Alpha platform In development, on the same ligands ANTIGEN PEPTIDE LIGAND LINKER CHELATOR RADIONUCLIDE under 40 residues · binds one antigen holds the metal for the life of the dose one ligand, three payloads

Target

Antigens over-expressed on solid tumors with few treatment options and expressed at low levels in healthy adult tissue. Because the same ligand is used for imaging, the presence of the target is confirmed in each patient by a PET scan (positron emission tomography) before any therapy is given.

Ligand

Peptides of fewer than 40 amino acids. Molecules of this size penetrate tumor tissue and are cleared from the blood within hours, so the dose to healthy organs is small in comparison to that from an antibody, which circulates for days. Each candidate is designed against a defined patch on the target and ranked on what is measured rather than on what is predicted.

Radionuclide

Gallium-68 (⁶⁸Ga) for PET imaging and lutetium-177 (¹⁷⁷Lu) for therapy, on the same ligand. Since ¹⁷⁷Lu also emits gamma rays, the dose delivered to the tumor and to each organ is measured by SPECT after each treatment rather than assumed. An alpha-emitting payload is in development for the tumors that need more energy deposited over a shorter range.

Discovery

A radioligand needs a binder with a long residence time on the target, a short residence time everywhere else, and a site at which a chelator can be attached without disturbing either. No single discovery technology delivers all three on every target, so we run several in parallel and let the measured data decide which candidates go forward. Peptides are the first modality, since a peptide of fewer than 40 residues clears within hours and carries a chelator without difficulty; small molecules are pursued where the target has a pocket a peptide cannot reach.

Peptide discovery

  • Display technology. Phage and mRNA display libraries of 10⁹ or more cyclic peptides, panned against the recombinant target and against cells which express it.
  • High-throughput screening of synthetic peptide libraries. Arrayed libraries made on resin, screened for binding and for the off-rate directly.
  • Tag-less peptide screening. Label-free methods (surface plasmon resonance and grating-coupled interferometry) which measure the native peptide rather than a tagged derivative.
  • Protein-inspired design. Binding loops and interfaces taken from the target's natural partners and constrained into a macrocycle.
  • Virtual screening. Docking and co-folding of large virtual libraries against a defined patch on the target, before anything is synthesized.
  • Computational and generative design. Structure-based generation of new binders against the patch, gated by the limits the radiochemistry imposes.

Small-molecule discovery

  • High-throughput screening. Biochemical and cell-based screens of diverse compound collections against the target.
  • Fragment-based discovery. Low-molecular-weight fragments found by biophysical methods and grown or linked into leads.
  • DNA-encoded libraries. Selections across libraries of 10⁸ or more compounds, each carrying a DNA barcode which identifies it.
  • Structure- and computation-led design. Design from the crystal or predicted structure of the target, with the chelator attachment point fixed from the start.
  • Phenotypic screening. Screens read out on the behavior of the tumor cell rather than on a single purified protein.
  • Generative chemistry. Machine-learned generation and scoring of new chemotypes within the property space a radioligand requires.
Selection

Whichever technology finds the binder, the same measurements decide whether it is a candidate: off-rate on the target, clearance from healthy organs, and a labeling site that survives conjugation.

Optimization and Radiochemistry

A hit becomes a candidate by surviving three stages in order: lead optimization, candidate nomination, and radiolabeling method development. Each stage has its own measurements and its own reasons for stopping a compound, and the radiochemistry is in the room from the first of them, since a binder which cannot be labeled is not a drug.

Lead optimization

Affinity is the entry ticket; residence time, selectivity, and clearance are what are optimized. Analogs are made by solid-phase synthesis, measured label-free for the off-rate, and profiled for stability in serum, for organ clearance, and for the net charge and lipophilicity that govern where the dose goes. The chelator and its attachment site are fixed early, because moving them later changes everything measured before.

Candidate nomination

The ⁶⁸Ga and ¹⁷⁷Lu versions of the two or three best compounds are compared head to head in tumor-bearing animals for tumor uptake, retention, and organ clearance. A candidate is nominated on that data, with a written target product profile and the first specification of what the clinical product will have to be.

Radiolabeling method development

The labeling reaction is developed into a method a manufacturing site can run: precursor amount, buffer, temperature and time, radiolytic protection, and the quality-control methods which prove radiochemical purity and identity. The method is written so that the same procedure serves the imaging agent and the therapy, and so that it transfers without rework.

Radionuclides

Two radionuclides carry every program into the clinic, one for imaging and one for therapy, and an alpha-emitting payload is in development on the same ligands. Each ligand carries a chelator that binds ⁶⁸Ga for PET and ¹⁷⁷Lu for therapy, and the imaging agent enters the clinic first, to show that the target is present in patients and that the ligand reaches it at a useful concentration.

Imaging

Gallium-68

PET

Beta therapy

Lutetium-177

β⁻, γ for SPECT

Alpha therapy

Alpha emitter

in development

RadionuclideEmissionHalf-lifeRange in tissueRoleSupply
Gallium-68β⁺ (positron), for PET68 minNot used for therapyImaging and patient selection⁶⁸Ge/⁶⁸Ga generator at the imaging site
Lutetium-177β⁻, with γ lines at 113 and 208 keV for SPECT6.65 d0.67 mm mean, 2 mm maximumTherapy, with post-treatment imaging and dosimetryReactor-produced, no-carrier-added
Alpha emitterα, one or more per decayHours to days50–100 µmAlpha therapy, in developmentUndisclosed

Half-lives and emissions are evaluated values (ENSDF, IAEA). Ranges are approximate values in soft tissue.

Alpha and Beta Emitters

Beta and alpha particles differ in energy, in range, and in the kind of damage they do to DNA, and each suits a different kind of tumor. Beta emitters such as ¹⁷⁷Lu are suited to larger and heterogeneous tumors, since a beta particle travels far enough to irradiate neighboring cells that carry less of the target (the cross-fire effect). Alpha emitters are suited to small metastases and to resistant, hypoxic cells, since a few alpha traversals kill a cell regardless of its oxygen supply. Our ligands are designed to carry either, which is why the chelator and the conjugation site are fixed before a panel is synthesized.

decay one cell ≈ 15 µm α particle 50–100 µm, about 3 to 7 cells β⁻ particle, mean 0.67 mm, about 45 cells (¹⁷⁷Lu) β⁻ particle, maximum 2 mm, about 130 cells (¹⁷⁷Lu) 00.5 mm1.0 mm1.5 mm2.0 mm DISTANCE FROM THE DECAY, TO ONE SCALE
An alpha particle stops within a few cells of the cell which bound the ligand. A lutetium-177 beta particle reaches dozens of cells on average, and the most energetic reach several hundred.
Energy
Alpha5–9 MeV per particle
Beta0.1–2.3 MeV; lutetium-177 up to 0.5 MeV
Range
Alpha50–100 µm, a few cell diameters
Beta0.5–12 mm, dozens to thousands of cells
Ionization density
AlphaAbout 80–100 keV per µm
BetaAbout 0.2 keV per µm
DNA damage
AlphaClustered double-strand breaks, independent of oxygen
BetaMostly single-strand breaks, dependent on oxygen
Hits to kill a cell
AlphaA handful of traversals
BetaThousands

Manufacturing

A radiopharmaceutical is made for a named patient and used within hours of manufacture, so there is no opportunity to test a batch at length before it is given. Quality has to be designed into the process from the first clinical batch, the process has to transfer to a site that can run it on the day it is needed, and the whole supply chain has to work to the clock set by the half-life. CMC is a core competence of the company, not a service bought in.

Method development and validation

Analytical methods for identity, radiochemical and radionuclidic purity, specific activity, and sterility are developed and validated to ICH and pharmacopoeial standards, with acceptance criteria set by the product's own data rather than borrowed from another.

Tech transfer and CDMO management

Processes and methods are transferred to contract manufacturers with complete documentation, on-site training, and comparability data, and the sites are managed under quality agreements that state who decides what. We have done this on both sides of the agreement.

Just-in-time manufacturing

Every dose is scheduled backwards from the time of administration: radionuclide delivery, labeling, release testing, and transport are planned per patient with the decay accounted for at each step, so that the activity that arrives is the activity that was prescribed.

Quality control and assurance

Release testing that fits inside the half-life, a stability program that sets the shelf life, and a quality system that keeps the whole process in agreement with the approved filing.

Timing

Every dose is manufactured, released, and administered on a schedule set by the half-life of the radionuclide, which for ⁶⁸Ga is 68 minutes. The process has to work the first time, since there is rarely time to repeat it.